Pyrrolo-, pyrazolo-, imidazo-pyrimidine and pyridine compounds that inhibit mnk1 and mnk2

ABSTRACT

The present invention provides synthesis, pharmaceutically acceptable formulations and uses of compounds in accordance with Formula I. or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof. 
     
       
         
         
             
             
         
       
     
     For Formula I compounds A 1 , A 2 . A 3 , A 4 , A 5 , A 6 , A 7 , W 1 , R 1 , R 2 , R 3 , R 4 , R 5a , R 5b , R 6 , R 7 , R 7a , R 7b , R 8 , R 8a , R 8b , R 9 , R 9a , R 9b  and R 10  and subscripts “m” and “n” are as defined in the specification. The inventive Formula I compounds are inhibitors of Mnk and find utility in any number of therapeutic applications, including but not limited to treatment of inflammation and various cancers.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication No. 62/247,966, filed Oct. 29, 2015, which is incorporatedherein by reference in its entirety.

FIELD

The present invention generally relates to compounds having activity asinhibitors of MAP kinase-interacting kinase (Mnk), for example Mnk1 andMnk2, as well as to related compositions and methods for utilizing theinventive compounds as therapeutic agents for treatment of Mnk dependentdiseases, including the treatment of cancer.

BACKGROUND

Eukaryotic initiation factor 4E (eIF4E) is a general translation factorbut it has the potential to enhance preferentially the translation ofmessenger RNAs (mRNAs) that lead to production of malignancy-associatedproteins. This selectivity may relate to an increased requirement foreIF4E and its binding partners for the translation of mRNAs containingextensive secondary structure in their 5′-untranslated regions(5′-UTRs). These mRNAs include those encoding certain proteins thatcontrol cell cycle progression and tumorigenesis. Under normal cellularconditions the translation of these malignancy-associated mRNAs issuppressed as the availability of active eIF4E is limited; however,their levels can increase when eIF4E is over-expressed orhyperactivated. Elevated levels of eIF4E have been found in many typesof tumors and cancer cell lines including cancers of the colon, breast,bladder, lung, prostate, gastrointestinal tract, head and neck.Hodgkin's lymphomas and neuroblastomas.

Initiation of cap-dependent translation is thought to depend on theassembly of eIF4F, an initiation factor complex including eIF4E, thescaffold protein eIF4G, and the RNA helicase eIF4A. Because eIF4E is theonly one of these proteins that binds directly to the mRNA capstructure, it is the key factor for the assembly of eIF4F at the 5′ cap.The scaffold protein, eIF4G, also recruits the 40S ribosomal subunit tothe mRNA via its interaction with eIF3 and binds eIF4B, a protein thataids the RNA-helicase function of eIF4A, thus facilitating thetranslation of mRNAs that contain structured 5′-UTRs. The availabilityof eIF4E as part of the eIF4F complex is a limiting factor incontrolling the rate of translation, and therefore eIF4E is an importantregulator of mRNA translation.

Regulation of eIF4E activity forms a node of convergence of thePI3K/Akt/mTOR and Ras/Raf/MAPK signaling pathways. The PI3K(phosphoinositide 3-kinase)/PTEN (phosphatase and tensin homologuedeleted on chromosome ten)/Akt/mTOR (mammalian target of rapamycin)pathway is often involved in tumorgenesis and in sensitivity andresistance to cancer therapy.

Deregulated signaling through the PI3K/PTEN/Akt/mTOR pathway is oftenthe result of genetic alterations in critical components of this pathwayand/or mutations at upstream growth factor receptors or signalingcomponents. PI3K initiates a cascade of events when activated by, forexample, extracellular growth factors, mitogens, cytokines and/orreceptors, PDK1 activates Akt, which in turn phosphorylates andinactivates the tumor suppressor complex comprising TSC1 and 2 (tuberoussclerosis complex 1/2), resulting in the activation of mTORC1 (target ofrapamycin complex 1) by Rheb-GTP. Activation of PDK1 and Akt by PI3Ks isnegatively regulated by PTEN.

PTEN is a critical tumor suppressor gene and is often mutated orsilenced in human cancers. Its loss results in activation of Akt andincreases downstream mTORC1 signaling. The involvement of mTOR complex 1(mTORC1) in neoplastic transformation appears to depend on itsregulatory role toward the eIF4F complex; overexpression of eIF4E canconfer resistance to rapamycin, mTORC regulates the eIF4F complexassembly that is critical for the translation of mRNAs associated withcell growth, prevention of apoptosis and transformation, mTORC1 achievesthis by phosphorylation and inactivation of 4E-BPs and the subsequentdissociation of 4E-BPs from eIF4E. This then enables eIF4E to interactwith the scaffold protein eIF4G, permitting assembly of the eIF4Fcomplex for the translation of structured mRNAs, mTORC1 also promotesactivation of the translational activator, S6K, which phosphorylates theribosomal protein S6 and other substrates, including eIF4B, mTORC1signaling is inhibited by rapamycin and its analogues (rapalogs),although these compounds act allosterically, rather than directlyinhibiting mTOR kinase activity.

Given the importance of the PI3K/Akt/mTOR pathway in regulating mRNAtranslation of genes that encode for pro-oncogenic proteins andactivated mTORC1 signaling in a high proportion of cancers, thesekinases have been actively pursued as oncology drug targets. A number ofpharmacological inhibitors have been identified, some of which havereached advanced clinical stages. However, it has recently become clearthat the mTOR pathway participates in a complicated feedback loop thatcan impair activation of Akt. It has been shown that prolonged treatmentof cancer cells or patients with mTOR inhibitors causes elevated PI3Kactivity that leads to phosphorylation of Akt and eIF4E, and promotescancer cell survival, eIF4E, acting downstream of Akt and mTOR,recapitulates Akt's action in tumorigenesis and drug resistance, and Aktsignaling via eIF4E is an important mechanism of oncogenesis and drugresistance in vivo.

In addition to the PI3K/Akt/mTOR pathway, eIF4E is also the target ofthe Ras/Raf/MAP signaling cascade which is activated by growth factorsand for the stress-activated p38 MAP kinase pathway. Erk1/2 and p38 thenphosphorylate MAP kinase-interacting kinase 1 (Mnk1) and MAPkinase-interacting kinase 2 (Mnk2). The Erk pathway is also activated inmany cancers, reflecting, for example, activating mutations in Ras(found in around 20% of tumors) or loss of function of the RasGTPase-activator protein NF1. Mnk1 and Mnk2 are threonine/serine proteinkinases and specifically phosphorylate serine 209 (Ser209) of eIF4Ewithin the cIF4F complex, by virtue of the interaction between eIF4E andthe Mnks, which serves to recruit Mnks to act on eIF4E. Mice withmutated eIF4E, in which Scr209 is replaced by alanine, shows no eIF4Ephosphorylation and significantly attenuated tumor growth.Significantly, while Mnk activity is necessary for eIF4E-mediatedoncogenic transformation, it is dispensable for normal development.Pharmacologically inhibiting Mnks thus presents an attractivetherapeutic strategy for cancer.

Despite increased understanding of Mnk structure and function, littleprogress has been made with regard to the discovery of pharmacologicalMnk inhibitors and relatively few Mnk inhibitors have been reported:CGP052088 (Tschopp et al., Mol Cell Biol Res Commun. 3(4):205-211,2000); CGP57380 (Rowlett et al., Am J Physiol Gastrointest LiverPhysiol. 294(2):G452-459, 2008); and Cercosporamide (Konicek et al.,Cancer Res. 71(5):1849-1857, 2011). These compounds, however, havemainly been used for the purpose of Mnk target validation. Morerecently, investigators have proposed further compounds for treatingdiseases influenced by the inhibition of kinase activity of Mnk1 and/orMnk2, including, for example, the compounds disclosed in WO 2014/044691and the various patent documents cited therein and the4-(dihydropyridinon-3-yl)amino-5-methylthieno[2,3,-d]pyrimidinesdisclosed by Yu et al., European Journal of Med. Chem., 95: 116-126,2015).

Accordingly, while advances have been made in this field there remains asignificant need in the art for compounds that specifically inhibit Mnkkinase activity, particularly with regard to Mnk's role in regulation ofcancer pathways, as well as for associated composition and methods. Thepresent invention fulfills this need and provides further relatedadvantages.

SUMMARY

The present invention is directed to compounds that inhibit or modulatethe activity of Mnk, as well as stereoisomers, tautomers andpharmaceutically acceptable salts of such compounds. The invention alsois directed to pharmaceutically acceptable compositions containing suchcompounds and associated methods for treating conditions that wouldbenefit from Mnk inhibition, such as cancer.

In one embodiment the invention is directed to compounds according toFormula I as well as to a stereoisomer, tautomer or pharmaceuticallyacceptable salt of such compounds, wherein

A¹ and A² independently are —N— or —CR^(8a):

A³ is —N— or —CR⁶;

A⁴ is —N— or —CR^(5b);

A⁵ is —NR⁷ or —CR^(7a)R^(7b);

A⁶ and A⁷ are independently —N— or —CR^(8a) when -------- represents abond, otherwise A⁶ and A⁷ are independently —NR⁵ or —CR⁸R^(8b);

W¹ is O, S, NH, NO(R⁹) or CR^(8a)R^(8b);

m and n independently are 1, 2 or 3;

R¹ and R² independently are —H, —NHR¹⁰, NHR¹⁰-alkylene, (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, cycloalkyl, heterocyclyl, heteroaryl,aryl, arylalkylene, cycloalkylalkylene, heterocyclylalkylene orheteroarylalkylene; or

R¹ and R² together with the carbon atom to which they are attached forma cycloalkyl or heterocyclyl ring;

R³ and R⁴ independently are —H, —OH, —CN, —SR¹⁰, S(O)₂(C₁-C₈) alkyl,—C(O)NHR¹⁰, —C(O)NR¹⁰R¹⁰, —NHR¹⁰, —NR¹⁰R¹⁰, NHR¹⁰-alkylene,NR¹⁰R¹⁰-alkylene, (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl.(C₁-C₈)haloalkyl, —O(C₁-C₈)alkyl, —O(C₁-C₈)haloalkyl,—O(C₁-C₈)alkyleneNHR¹⁰, —O(C₁-C₈)alkyleneNR¹⁰R¹⁰, cycloalkyl,heterocyclyl, heteroaryl, aryl, arylalkylene, cycloalkylalkylene,heterocyclylalkylene, heteroarylalkylene, alkylaminyl,alkylcarbonylaminyl, cycloalkylcarbonylaminyl, cycloalkylaminyl, orheterocyclylaminyl;

R^(5a) is —H, —OH, halogen, —CN, acetyl, —(C₁-C₈)alkyl, —S(C₁-C₈)alkyl,—(C₂-C₈)alkenyl, —(C₂-C₈)alkynyl, —O(C₁-C₈)alkyl, (C₁-C₈)haloalkyl,—NHR¹⁰, —NR¹⁰R¹⁰, —NHR¹⁰-alkylene, NR¹⁰R¹⁰-alkylene, or—O(C₁-C₈)haloalkyl;

R^(5b) and R⁶ is —H, —OH, —SH, —CN, —S(O)₂R¹⁰, halogen, —S(C₁-C₈)alkyl,—NHR¹⁰, —NR¹⁰R¹⁰, (C₁-C₈)alkyl, (C₂-C₈)alkenyl. (C₂-C₈)alkynyl,(C₁-C₈)haloalkyl, —O(C₁-C₈)haloalkyl, —O(C₁-C₈)alkyl,—O(C₁-C₈)alkyleneNHR¹⁰, —O(C₁-C₈)alkyleneNR¹⁰R¹⁰—(C₁-C₈)alkyleneNHR¹⁰,—(C₁-C₈)alkyleneNR¹⁰R¹⁰, —S(C₁-C₈)alkyl, cycloalkyl, heterocyclyl,heteroaryl, or aryl;

R⁷ is —H, —OH, acetyl, —(C₁-C₈)alkyl, —C(O)alkyl, —C(O)cycloalkyl,—C(O)O—(C₁-C₈)alkyl, cycloalkyl, aryl, heteroaryl or heterocyclyl;

R^(8a) and R^(8b) independently are —H, —OH, acetyl, —(C₁-C₈)alkyl,—O(C₁-C₈)alkyl, —C(O)alkyl, —C(O)cycloalkyl, —C(O)O—(C₁-C₈)alkyl,cycloalkyl, aryl, heteroaryl or heterocyclyl;

R⁸ is —H, —OH, acetyl. (C₁-C₈)alkyl, cycloalkyl, heterocyclyl,heteroaryl or aryl;

R^(8a) and R^(8b) independently are —H, —OH, —CN, acetyl, —SH,—S(O)₂R¹⁰, halogen, —S(C₁-C₈)alkyl, —NHR¹⁰, —NR¹⁰R¹⁰, (C₁-C₈)alkyl,(C₁-C₈)haloalkyl, —O(C₁-C₈)alkyl, —O(C₁-C₈)alkylNHR¹⁰,—O(C₁-C₈)alkylNR¹⁰R¹⁰, —(C₁-C₈)alkylNHR¹⁰, —(C₁-C₈)alkylNR¹⁰R¹⁰,cycloalkyl, heterocyclyl, heteroaryl or aryl:

R⁹, R^(9a) and R^(9b) are independently —H, (C₁-C₈)alkyl,(C₂-C₈)alkenyl. (C₂-C₈)alkynyl, cycloalkyl, heterocyclyl, heteroaryl,aryl, arylalkylene, cycloalkylalkylene, heterocyclylalkylene, orheteroarylalkylene, or

R^(9a) and R^(9b) together with the carbon atom to which they areattached form a cycloalkyl or heterocyclyl ring;

R¹⁰ is —H, —OH, —C(O)O(C₁-C₈)alkyl, —C(O)(C₁-C₈)alkyl, —C(O)—NH₂,—C(O)—NH(C₁-C₈)alkyl. NH₂—C(O)-alkylene, —S(C₁-C₈)alkyl, acetyl,—(C₁-C₈)alkyl, (C₂-C₈)alkenyl. (C₂-C₈)alkynyl, —O(C₁-C₈)alkyl, (C₁-C₈)haloalkyl, alkylcarbonylaminyl, alkylaminyl, —C(O)alkyl,—C(O)cycloalkyl, —C(O)O—(C₁-C₈)alkyl, aryl, heteroaryl, heterocyclyl orcycloalkyl;

wherein any alkyl, cycloalkyl, heterocyclyl, heteroaryl, aryl,arylalkylene, cycloalkylalkylene, heterocyclylalkylene,heteroarylalkylene, alkylaminyl, alkylcarbonylaminyl,cycloalkylcarbonylaminyl, cycloalkylaminyl or heterocyclylaminyl isoptionally substituted with 1, 2 or 3 groups selected from —OH, —CN,—SH, —S(O)NH₂, —S(O)NH₂, halogen. —NH₂, —NH(C₁-C₄)alkyl,—N[(C₁-C₄)alkyl]₂, —C(O)NH₂, —COOH, —COOMe, acetyl. —(C₁-C₈)alkyl,—O(C₁-C₈)alkyl (C₂-C₈)alkenyl. (C₂-C₈)alkynyl, haloalkyl, thioalkyl,cyanomethylene, alkylaminyl, NH₂—C(O)-alkylene, NH₂—C(O)-alkylene,—NH(Me)-C(O)-alkylene, —CH₂—C(O)-lower alkyl, —C(O)-lower alkyl,alkylcarbonylaminyl, cycloalkyl, cycloalkylalkylene,cycloalkylalkenylene, cycloalkylcarbonylaminyl, cycloalkylaminyl,—CH₂—C(O)-cycloalkyl, —C(O)-cycloalkyl, —CH₂—C(O)-aryl, —CH₂-aryl,—C(O)-aryl, —CH₂—C(O)-heterocycloalkyl, —C(O)-heterocycloalkyl,heterocyclylaminyl or heterocyclyl; and

--- represents the option of having a double bond.

The present invention also provides a pharmaceutical compositioncomprising (i) a therapeutically effective amount of at least onecompound according to Formula I or a stereoisomer, a tautomer or apharmaceutically acceptable salt thereof: (ii) in combination with apharmaceutically acceptable carrier, diluent or excipient.

Also provided by the present invention is a method for attenuating orinhibiting the activity of MnK in at least one cell overexpressing Mnk,comprising contacting the at least one cell with a compound according toclaim 1 or a stereoisomer, tautomer or pharmaceutically acceptable saltthereof.

According to the inventive method at least one cell is a colon cancercell, a gastric cancer cell, a thyroid cancer cell, a lung cancer cell,a leukemia cell, a B-cell lymphoma, a T-cell lymphoma, a hairy celllymphoma, Hodgkin's lymphoma cell, non-Hodgkin's lymphoma cell,Burkitt's lymphoma cell, a pancreatic cancer cell, a melanoma cell, amultiple melanoma cell, a brain cancer cell, a CNS cancer cell, a renalcancer cell, a prostate cancer cell, an ovarian cancer cell, or a breastcancer cell.

According to yet another embodiment the invention provides a method fortreating a Mnk dependent condition in a mammal in need thereofcomprising administering to the mammal (i) a therapeutically effectiveamount of at least one compound according to claim 1 or a stereoisomer,tautomer or pharmaceutically acceptable salt thereof, or (ii) apharmaceutical composition in accordance with the invention.

Compounds and pharmaceutically acceptable formulations in accordancewith the invention are useful for treating an Mnk dependent conditionsuch as colon cancer, gastric cancer, thyroid cancer, lung cancer,leukemia. B-cell lymphoma, T-cell lymphoma, hairy cell lymphoma.Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma,pancreatic cancer, melanoma, multiple melanoma, brain cancer. CNScancer, renal cancer, prostate cancer, ovarian cancer, or breast cancer.

The above embodiments and other aspects of the invention are readilyapparent in the detailed description that follows. Various referencesare set forth herein which describe in more detail certain backgroundinformation, procedures, compounds and/or compositions, and are eachhereby incorporated by reference in their entirety.

DETAILED DESCRIPTION

In the following description certain specific details are set forth inorder to provide a thorough understanding of various embodiments of theinvention. However, one skilled in the art will understand that theinvention may be practiced without these details. Unless the contextrequires otherwise, throughout the present specification and claims, theword “comprise” and variations thereof, such as, “comprises” and“comprising” are to be construed in an open, inclusive sense (i.e., as“including, but not limited to”).

Reference throughout this specification to “one embodiment” or “anembodiment” means that a particular feature, structure or characteristicdescribed in connection with the embodiment is included in at least oneembodiment of the present invention. Thus, the appearances of thephrases “in one embodiment” or “in an embodiment” in various placesthroughout this specification are not necessarily all referring to thesame embodiment. Furthermore, the particular features, structures, orcharacteristics may be combined in any suitable manner in one or moreembodiments.

Definitions

As used herein, and unless noted to the contrary, the following termsand phrases have the meaning noted below.

“Amino” refers to the —NH₂ substituent.

“Aminocarbonyl” refers to the —C(O)NH₂ substituent.

“Carboxyl” refers to the —CO₂H substituent.

“Carbonyl” refers to a —C(O)— or —C(═O)— group. Both notations are usedinterchangeably within the specification.

“Cyano” refers to the —C≡N substituent.

“Cyanoalkylene” refers to the -(alkylene)C═N substituent.

“Acetyl” refers to the —C(O)CH₃ substituent.

“Hydroxy” or “hydroxyl” refers to the —OH substituent.

“Hydroxyalkylene” refers to the -(alkylene)OH substituent.

“Oxo” refers to a ═O substituent.

“Thio” or “thiol” refer to a —SH substituent.

“Alkyl” refers to a saturated, straight or branched hydrocarbon chainradical consisting solely of carbon and hydrogen atoms, having from oneto twelve carbon atoms (C₁-C₁₂ alkyl), from one to eight carbon atoms(C₁-C₈ alkyl) or from one to six carbon atoms (C₁-C₆ alkyl), and whichis attached to the rest of the molecule by a single bond. Exemplaryalkyl groups include methyl, ethyl, n-propyl, 1-methylethyl(iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl),3-methylhexyl, 2-methylhexyl, and the like.

“Lower alkyl” has the same meaning as alkyl defined above but havingfrom one to four carbon atoms (C₁-C₄ alkyl).

“Alkenyl” refers to an unsaturated alkyl group having at least onedouble bond and from two to twelve carbon atoms (C₂-C₁₂ alkenyl), fromtwo to eight carbon atoms (C₂-C₈ alkenyl) or from two to six carbonatoms (C₂-C₆ alkenyl), and which is attached to the rest of the moleculeby a single bond, e.g., ethenyl, propenyl, butenyl, pentenyl, hexenyl,and the like.

“Alkynyl” refers to an unsaturated alkyl group having at least onetriple bond and from two to twelve carbon atoms (C₂-C₁₂ alkynyl), fromtwo to ten carbon atoms (C₂-C₁₀ alkynyl) from two to eight carbon atoms(C₂-C₈ alkynyl) or from two to six carbon atoms (C₂-C₆ alkynyl), andwhich is attached to the rest of the molecule by a single bond, e.g.,ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.

“Alkylene” or “alkylene chain” refers to a straight or branched divalenthydrocarbon (alkyl) chain linking the rest of the molecule to a radicalgroup, consisting solely of carbon and hydrogen, respectively. Alkylenescan have from one to twelve carbon atoms, e.g., methylene, ethylene,propylene, n-butylene, and the like. The alkylene chain is attached tothe rest of the molecule through a single or double bond. The points ofattachment of the alkylene chain to the rest of the molecule can bethrough one carbon or any two carbons within the chain. “Optionallysubstituted alkylene” refers to alkylene or substituted alkylene.

“Alkenylene” refers to divalent alkene. Examples of alkenylene includewithout limitation, ethenylene (—CH—CH—) and all stereoisomeric andconformational isomeric forms thereof. “Substituted alkenylene” refersto divalent substituted alkene. “Optionally substituted alkenylene”refers to alkenylene or substituted alkenylene.

“Alkynylene” refers to divalent alkyne. Examples of alkynylene includewithout limitation, ethynylene, propynylene. “Substituted alkynylene”refers to divalent substituted alkyne.

“Alkoxy” refers to a radical of the formula —OR_(a) where R_(a) is analkyl having the indicated number of carbon atoms as defined above.Examples of alkoxy groups include without limitation —O-methyl(methoxy), —O-ethyl (ethoxy), —O-propyl (propoxy), —O-isopropyl (isopropoxy) and the like.

“Alkylaminyl” refers to a radical of the formula —NHR_(a) or—NR_(a)R_(a) where each R_(a) is, independently, an alkyl radical havingthe indicated number of carbon atoms as defined above.

“Cycloalkylaminyl” refers to a radical of the formula —NHR_(a) whereR^(a) is a cycloalkyl radical as defined herein.

“Alkylcarbonylaminyl” refers to a radical of the formula —NHC(O)R^(a),where R_(a) is an alkyl radical having the indicated number of carbonatoms as defined herein.

“Cycloalkylcarbonylaminyl” refers to a radical of the formula—NHC(O)R_(a), where R_(a) is a cycloalkyl radical as defined herein.

“Alkylaminocarbonyl” refers to a radical of the formula —C(O)NHR_(a) or—C(O)NR_(a)R_(a), where each R_(a) is independently, an alkyl radicalhaving the indicated number of carbon atoms as defined herein.

“Cyclolkylaminocarbonyl” refers to a radical of the formula—C(O)NHR_(a), where R_(a) is a cycloalkyl radical as defined herein.

“Aryl” refers to a hydrocarbon ring system radical comprising hydrogen,6 to 18 carbon atoms and at least one aromatic ring. Exemplary aryls arehydrocarbon ring system radical comprising hydrogen and 6 to 9 carbonatoms and at least one aromatic ring; hydrocarbon ring system radicalcomprising hydrogen and 9 to 12 carbon atoms and at least one aromaticring; hydrocarbon ring system radical comprising hydrogen and 12 to 15carbon atoms and at least one aromatic ring; or hydrocarbon ring systemradical comprising hydrogen and 15 to 18 carbon atoms and at least onearomatic ring. For purposes of this invention, the aryl radical may be amonocyclic, bicyclic, tricyclic or tetracyclic ring system, which mayinclude fused or bridged ring systems. Aryl radicals include, but arenot limited to, aryl radicals derived from aceanthrylene,acenaphthylene, acephenanthrylene, anthracene, azulene, benzene,chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane,indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, andtriphenylene. “Optionally substituted aryl” refers to a aryl group or asubstituted aryl group.

“Arylene” denotes divalent aryl, and “substituted arylene” refers todivalent substituted aryl.

“Aralkyl” or “araalkylene” may be used interchangeably and refer to aradical of the formula —R_(b)—R_(c) where R_(b) is an alkylene chain asdefined herein and R_(c) is one or more aryl radicals as defined herein,for example, benzyl, diphenylmethyl and the like.

“Cycloalkyl” refers to a stable non-aromatic monocyclic or polycyclichydrocarbon radical consisting solely of carbon and hydrogen atoms,which may include fused or bridged ring systems, having from three tofifteen carbon atoms, preferably having from three to ten carbon atoms,three to nine carbon atoms, three to eight carbon atoms, three to sevencarbon atoms, three to six carbon atoms, three to five carbon atoms, aring with four carbon atoms, or a ring with three carbon atoms. Thecycloalkyl ring may be saturated or unsaturated and attached to the restof the molecule by a single bond. Monocyclic radicals include, forexample, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,and cyclooctyl. Polycyclic radicals include, for example, adamantyl,norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like.

“Cycloalkylalkylene” or “cycloalkylalkyl” may be used interchangeablyand refer to a radical of the formula —R_(b)R_(c) where R_(b) is analkylene chain as defined herein and R_(e) is a cycloalkyl radical asdefined herein. In certain embodiments. R_(b) is further substitutedwith a cycloalkyl group, such that the cycloalkylalkylene comprises twocycloalkyl moieties. Cyclopropylalkylene and cyclobutylalkylene areexemplary cycloalkylalkylene groups, comprising at least one cyclopropylor at least one cyclobutyl group, respectively.

“Fused” refers to any ring structure described herein which is fused toan existing ring structure in the compounds of the invention. When thefused ring is a heterocyclyl ring or a heteroaryl ring, any carbon atomon the existing ring structure which becomes part of the fusedheterocyclyl ring or the fused heteroaryl ring may be replaced with anitrogen atom.

“Halo” or “halogen” refers to bromo (bromine), chloro (chlorine), fluoro(fluorine), or iodo (iodine).

“Haloalkyl” refers to an alkyl radical having the indicated number ofcarbon atoms, as defined herein, wherein one or more hydrogen atoms ofthe alkyl group are substituted with a halogen (halo radicals), asdefined above. The halogen atoms can be the same or different.

Exemplary haloalkyls are trifluoromethyl, difluoromethyl,trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl,3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and the like.

“Heterocyclyl”, heterocycle”, or “heterocyclic ring” refers to a stable3- to 18-membered saturated or unsaturated radical which consists of twoto twelve carbon atoms and from one to six heteroatoms, for example, oneto five heteroatoms, one to four heteroatoms, one to three heteroatoms,or one to two heteroatoms selected from the group consisting ofnitrogen, oxygen and sulfur. Exemplary heterocycles include withoutlimitation stable 3-15 membered saturated or unsaturated radicals,stable 3-12 membered saturated or unsaturated radicals, stable 3-9membered saturated or unsaturated radicals, stable 8-membered saturatedor unsaturated radicals, stable 7-membered saturated or unsaturatedradicals, stable 6-membered saturated or unsaturated radicals, or stable5-membered saturated or unsaturated radicals.

Unless stated otherwise specifically in the specification, theheterocyclyl radical may be a monocyclic, bicyclic, tricyclic ortetracyclic ring system, which may include fused or bridged ringsystems; and the nitrogen, carbon or sulfur atoms in the heterocyclylradical may be optionally oxidized; the nitrogen atom may be optionallyquaternized; and the heterocyclyl radical may be partially or fullysaturated. Examples of non-aromatic heterocyclyl radicals include, butare not limited to, azetidinyl, dioxolanyl, thienyl[1.3]dithianyl,decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl,isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl,2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl,piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl,quinuclidinyl, thiazolidinyl, tetrahydrofuryl, thietanyl, trithianyl,tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl,1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Heterocyclylsinclude heteroaryls as defined herein, and examples of aromaticheterocyclyls are listed in the definition of heteroaryls below.

“Heterocyclylalkyl” or “heterocyclylalkylene” refers to a radical of theformula —R_(b)R_(f) where R_(b) is an alkylene chain as defined hereinand R_(f) is a heterocyclyl radical as defined above, and if theheterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl maybe attached to the alkyl radical at the nitrogen atom.

“Heteroaryl” or “heteroarylene” refers to a 5- to 14-membered ringsystem radical comprising hydrogen atoms, one to thirteen carbon atoms,one to six heteroatoms selected from the group consisting of nitrogen,oxygen and sulfur, and at least one aromatic ring. For purposes of thisinvention, the heteroaryl radical may be a stable 5-12 membered ring, astable 5-10 membered ring, a stable 5-9 membered ring, a stable 5-8membered ring, a stable 5-7 membered ring, or a stable 6 membered ringthat comprises at least 1 heteroatom, at least 2 heteroatoms, at least 3heteroatoms, at least 4 heteroatoms, at least 5 heteroatoms or at least6 heteroatoms.

Heteroaryls may be a monocyclic, bicyclic, tricyclic or tetracyclic ringsystem, which may include fused or bridged ring systems; and thenitrogen, carbon or sulfur atoms in the heteroaryl radical may beoptionally oxidized: the nitrogen atom may be optionally quaternized.The heteroatom may be a member of an aromatic or non-aromatic ring,provided at least one ring in the heteroaryl is aromatic. Examplesinclude, but are not limited to, azepinyl, acridinyl, benzimidazolyl,benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl,benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl,1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl,benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl,benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl,imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl,isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl,oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl,1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl.1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl,phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl,pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl,quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl,thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, andthiophenyl (i.e. thienyl).

“Heteroarylalkyl” or “heteroarylalkylene” refers to a radical of theformula —R_(b)R_(g) where R_(b) is an alkylene chain as defined aboveand R₈ is a heteroaryl radical as defined above.

“Thioalkyl” refers to a radical of the formula —SR_(a) where R_(a) is analkyl radical as defined above containing one to twelve carbon atoms, atleast 1-10 carbon atoms, at least 1-8 carbon atoms, at least 1-6 carbonatoms, or at least 1-4 carbon atoms.

“Heterocyclylaminyl” refers to a radical of the formula —NHR_(f) whereR_(f) is a heterocyclyl radical as defined above.

“Thione” refers to a ═S group attached to a carbon atom of a saturatedor unsaturated (C₃-C₈)cyclic or a (C₁-C₈)acyclic moiety.

“Sulfoxide” refers to a —S(O)— group in which the sulfur atom iscovalently attached to two carbon atoms.

“Sulfone” refers to a —S(O)₂— group in which a hexavalent sulfur isattached to each of the two oxygen atoms through double bonds and isfurther attached to two carbon atoms through single covalent bonds.

The term “oxime” refers to a —C(R_(a))N—OR_(a) radical where R_(a) ishydrogen, lower alkyl, an alkylene or arylene group as defined above.

The compound of the invention can exist in various isomeric forms, aswell as in one or more tautomeric forms, including both single tautomersand mixtures of tautomers. The term “isomer” is intended to encompassall isomeric forms of a compound of this invention, including tautomericforms of the compound.

Some compounds described here can have asymmetric centers and thereforeexist in different enantiomeric and diastereomeric forms. A compound ofthe invention can be in the form of an optical isomer or a diastereomer.Accordingly, the invention encompasses compounds of the invention andtheir uses as described herein in the form of their optical isomers,diastereoisomers and mixtures thereof, including a racemic mixture.Optical isomers of the compounds of the invention can be obtained byknown techniques such as asymmetric synthesis, chiral chromatography, orvia chemical separation of stereoisomers through the employment ofoptically active resolving agents.

Unless otherwise indicated “stereoisomer” means one stereoisomer of acompound that is substantially free of other stereoisomers of thatcompound. Thus, a stereomerically pure compound having one chiral centerwill be substantially free of the opposite enantiomer of the compound. Astereomerically pure compound having two chiral centers will besubstantially free of other diastereomers of the compound. A typicalstereomerically pure compound comprises greater than about 80% by weightof one stereoisomer of the compound and less than about 20% by weight ofother stereoisomers of the compound, for example greater than about 90%by weight of one stereoisomer of the compound and less than about 10% byweight of the other stereoisomers of the compound, or greater than about95% by weight of one stereoisomer of the compound and less than about 5%by weight of the other stereoisomers of the compound, or greater thanabout 97% by weight of one stereoisomer of the compound and less thanabout 3% by weight of the other stereoisomers of the compound.

If there is a discrepancy between a depicted structure and a name givento that structure, then the depicted structure controls. Additionally,if the stereochemistry of a structure or a portion of a structure is notindicated with, for example, bold or dashed lines, the structure orportion of the structure is to be interpreted as encompassing allstereoisomers of it. In some cases, however, where more than one chiralcenter exists, the structures and names may be represented as singleenantiomers to help describe the relative stereochemistry. Those skilledin the art of organic synthesis will know if the compounds are preparedas single enantiomers from the methods used to prepare them.

In this description a “pharmaceutically acceptable salt” is apharmaceutically acceptable, organic or inorganic acid or base salt of acompound of the invention. Representative pharmaceutically acceptablesalts include, e.g., alkali metal salts, alkali earth salts, ammoniumsalts, water-soluble and water-insoluble salts, such as the acetate,amsonate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate,benzonate, bicarbonate, bisulfate, bitartrate, borate, bromide,butyrate, calcium, calcium edetate, camsylate, carbonate, chloride,citrate, clavulariate, dihydrochloride, edetate, edisylate, estolate,esylate, fiunarate, gluceptate, gluconate, glutamate,glycollylarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine,hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate,lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate,methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate,N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate,oxalate, palmitate, pamoate (1,1-methene-bis-2-hydroxy-3-naphthoate,einbonate), pantothenate, phosphate/diphosphate, picrate,polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate,subacetate, succinate, sulfate, sulfosaliculate, suramate, tannate,tartrate, teoclate, tosylate, triethiodide, and valerate salts. Apharmaceutically acceptable salt can have more than one charged atom inits structure. In this instance the pharmaceutically acceptable salt canhave multiple counterions. Thus, a pharmaceutically acceptable salt canhave one or more charged atoms and/or one or more counterions.

The terms “treat”, “treating” and “treatment” refer to the ameliorationor eradication of a disease or symptoms associated with a disease. Incertain embodiments, such terms refer to minimizing the spread orworsening of the disease resulting from the administration of one ormore prophylactic or therapeutic agents to a patient with such adisease. In the context of the present invention the terms “treat”,“treating” and “treatment” also refer to:

(i) preventing the disease or condition from occurring in a mammal, inparticular, when such mammal is predisposed to the condition but has notyet been diagnosed as having it;(ii) inhibiting the disease or condition, i.e., arresting itsdevelopment;(iii) relieving the disease or condition, i.e., causing regression ofthe disease or condition; or(iv) relieving the symptoms resulting from the disease or condition,i.e., relieving pain without addressing the underlying disease orcondition. As used herein, the terms “disease” and “condition” may beused interchangeably or may be different in that the particular maladyor condition may not have a known causative agent (so that etiology hasnot yet been worked out) and it is therefore not yet recognized as adisease but only as an undesirable condition or syndrome, wherein a moreor less specific set of symptoms have been identified by clinicians.

The term “effective amount” refers to an amount of a compound of theinvention or other active ingredient sufficient to provide a therapeuticor prophylactic benefit in the treatment or prevention of a disease orto delay or minimize symptoms associated with a disease. Further, atherapeutically effective amount with respect to a compound of theinvention means that amount of therapeutic agent alone, or incombination with other therapies, that provides a therapeutic benefit inthe treatment or prevention of a disease. Used in connection with acompound of the invention, the term can encompass an amount thatimproves overall therapy, reduces or avoids symptoms or causes ofdisease, or enhances the therapeutic efficacy or synergies with anothertherapeutic agent.

The terms “modulate”, “modulation” and the like refer to the ability ofa compound to increase or decrease the function, or activity of, forexample, MAP kinase interacting kinase (Mnk). “Modulation”, in itsvarious forms, is intended to encompass inhibition, antagonism, partialantagonism, activation, agonism and/or partial agonism of the activityassociated with Mnk. Mnk inhibitors are compounds that bind to,partially or totally block stimulation, decrease, prevent, delayactivation, inactivate, desensitize, or down regulate signaltransduction. The ability of a compound to modulate Mnk activity can bedemonstrated in an enzymatic assay or a cell-based assay.

A “patient” or subject” includes an animal, such as a human, cow, horse,sheep, lamb, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbitor guinea pig. The animal can be a mammal such as a non-primate and aprimate (e.g., monkey and human). In one embodiment, a patient is ahuman, such as a human infant, child, adolescent or adult.

The term “prodrug” refers to a precursor of a drug, a compound whichupon administration to a patient, must undergo chemical conversion bymetabolic processes before becoming an active pharmacological agent.Exemplary prodrugs of compounds in accordance with Formula I are esters,acetamides, and amides.

Compounds of the Invention

The present invention generally is directed to compounds encompassed bythe genus of Formula I, a stereoisomer, a tautomer or a pharmaceuticallyacceptable salt thereof.

For Formula I compounds A¹, A². A³, A⁴, A⁵, A⁶, A⁷, W¹, R¹, R², R³, R⁴,R^(5a), R^(5b), R⁶, R⁷, R^(7a), R^(7b), R⁸, R^(8a), R^(8b), R⁹, R^(9a),R^(9b) and R¹⁰ and subscripts “m” and “n” are as defined in thespecification. Described below are specific embodiments of Formula Icompounds.

In one embodiment A¹ and A² are —CR^(5a).

In another embodiment A¹ is —N— and A² is —CH or —C(Me). In yet anotherembodiment A¹ is —CH and A² is —N—.

In one embodiment A³ is —CR⁶.

In one embodiment A⁴ is —CR⁵¹.

In one embodiment A⁵ is an —NR⁷.

In another embodiment A⁵ is —CR^(7a)R^(7b).

In one embodiment A⁶ and A⁷ are —CR^(8a).

In one embodiment W¹ is O.

In one embodiment subscript “m” and subscript “n” are 1.

In another embodiment subscript “m” is 2 and subscript “n” is 1. In yetanother embodiment subscripts “m” and “n” are both 2.

In one embodiment R¹ and R² independently are —H, (C₁-C₈)alkyl, —NHR¹⁰or NHR¹⁰-alkylene.

In one embodiment R¹ and R² are —H, —NH₂, —NH(Me), —N(Me)₂, methyl,ethyl, propyl, isopropyl, butyl, sec-butyl, I-butyl, isobutyl, pentyl,hexyl, methylene-NH[C(O)OMe] or ethylene-NH[C(O)OMe].

In one embodiment at least one of R¹ or R² is a halogen substituted(C₁-C₈)alkyl, (C₂-C₈)alkenyl. (C₂-C₈)alkynyl, cycloalkyl, heterocyclyl,heteroaryl, aryl, arylalkylene, cycloalkylalkylene, heterocyclylalkyleneor heteroarylalkylene.

In one embodiment R¹ and R² together with the carbon atom to which theyare attached form a cycloalkyl ring. In an embodiment the cycloalkylring is cyclobutyl, cyclopentyl, cyclohexyl, 2,2-dimethylcyclobutyl,4-aminocyclohexyl, 4-methylcyclohexyl, 4-ethylcyclohexyl,2,2-difluoroethyl-4-cyclohexyl, 4,4-difluorocyclohexy,4-cyanocyclohexyl, 4-trifluoromethylcyclohexyl, 4-hydroxycyclohexyl,3-hydroxycyclopently, 3-aminocyclopentyl or a 3-methylcyclopentyl ringsystem. In yet another embodiment the cycloalkyl ring is cyclobutyl,cyclopentyl or cyclohexyl.

In one embodiment R¹ and R² together with the carbon atom to which theyare attached form a heterocyclyl ring. In an embodiment the heterocyclylring is 1-(2,2-difluoroethyl)piperidine or 1-methylpiperidine.

In one embodiment R³, R⁴, and R^(5b) independently are—O(C₁-C₈)alkyleneNHR¹⁰ or —O(C₁-C₈)alkyleneNR¹⁰R¹⁰. In anotherembodiment R³, R⁴, and R^(5b) independently are —O(CH₂)NH₂, —O(CH₂)₂NH₂,—O(CH₂)₃NH₂, —O(CH₂)NH(Me), —O(CH₂)₂NH(Me) or —O(CH₂)₃NH(Me). In yetanother embodiment R³, R⁴, and R-5 independently are —O(CH₂)N(Me)₂,—O(CH₂)₂N(Me)₂ or —O(CH₂)₃N(Me)₂.

In one embodiment R³, R⁴, and R^(5b) independently are —H, —OH, CN,—C(O)NH₂, —C(O)NH(Me), —NH₂, —NH(Me), —N(Me)₂, —NH₂-methylene,—NH₂-ethylene, methyl, ethyl, propyl, n-butyl, i-butyl, t-butyl, hexyl,methoxy, ethoxy, propoxy, butoxy, chloromethyl, fluoromethyl,dichloromethyl, chlorofluoromethyl, trifluoromethyl, chloroethyl,1,2-dichloroethyl or chlorofluoroethyl.

In one embodiment R^(5a) is —H, —OH, halogen, —CN, acetyl or—(C₁-C₈)alkyl. In another embodiment R^(5a) is methyl, ethyl, propyl orbutyl.

In one embodiment R⁶ is amino, methylamino, —CN, —O(C₁-C₈)alkyleneNHR¹⁰,−O(C₁-C₈)alkyleneNR¹⁰R¹⁰, —(C₁-C₈)alkyleneNHR¹⁰ or—(C₁-C₈)alkyleneNR¹⁰R¹⁰.

In another embodiment R⁶ is —H, —OH, chlorine, fluorine, methyl ethyl,propyl and the like.

In one embodiment R⁷, R^(7a) and R^(7b) are hydrogen.

In one embodiment R⁸, R^(8a) and R^(8b) independently are —H,heterocyclyl, heteroaryl or aryl. In another embodiment R⁸, R^(8a) andR^(8b) independently are pyridine, 1-(2,2-difluoroethyl)piperidine,1-difluoromethyl piperidine, N-methylpyrazole, thioimidazole, piperidineor N-methylpiperidine, phenyl, 2-chlorophenyl, 3-chlorophenyl,4-chlorophenyl, 2-cyanophenyl, 3-cyanophenyl or 4-cyanophenyl.

In one embodiment R^(8a) and R^(8b) independently are —H, —OH, —CN, Cl,F, methyl, ethyl propyl, chloromethyl, fluoromethyl, chlorofluoromethyl,—NH(Me) or —N(Me)₂.

In one embodiment R⁹, R^(9a) and R^(9b) are independently —H or—(C₁-C₈)alkyl.

In one embodiment R¹⁰ is —H, —OH, methyl, ethyl, propyl, butyl, t-butyl,acetyl. —COOMe, —NH₂, —NH(Me), or —N(Me)₂.

In one embodiment A¹ is —N, A², A³, A⁴, A⁶, and A⁷ are —CH, A⁵ is —NH,W¹ is O, and subscripts “m” and “n” are both 1.

In another embodiment A¹ is —N, A² is —CH, A³ is —C(Cl), —C(F), —C(Me)or —C(OH), A⁴. A⁶, and A⁷ are —CH, A⁵ is —NH, W¹ is O, and subscripts“m” and “n” are both 1.

In another embodiment A¹ is —N, A² is —CH, A³ is —C(alkyl) or—C(halogen). A⁴ is —CH, A⁵ is —NH. A⁶ and A⁷ are both —CR^(8a), W¹ is O,and subscripts “m” and “n” are both 1. In an embodiment —CR^(8a) is—C(pyridyl), —C(N-methylpyrazole), —C(2-chlorophenyl) or—C(2-cyanophenyl).

In another embodiment A¹ is —N, A², A³, A⁴ are —CH, A⁵ is —NH, A⁶ and A⁷are —N—, W¹ is O, and subscripts “m” and “n” are both 1.

In another embodiment one of A⁶ or A⁷ is —N and the other of A⁶ or A⁷ is—CH, —C(pyridyl), —C(N-methylpyrazole), —C(2-chlorophenyl) or—C(2-cyanophenyl).

In another embodiment A¹ is —N, A², A³, A⁴ are —CH, A⁵ is —NH, A⁶ and A⁷independently are CH₂— or —CH(Me). W¹ is O, and subscripts “m” and “n”are both 1.

In another embodiment one of A⁶ or A⁷ is —CH₂ or —CH(Me) and the otherof A⁶ or A⁷ is —NH.

In one embodiment subscript “m” is 2 and subscript “n” is 1, A¹ is —N,A², A³ and A⁴ are —CH, A⁵ is —NH and A⁶ and A⁷ are —CH₂.

The inventive compounds according to Formula I may beisotopically-labeled by having one or more atoms replaced by an atomhaving a different atomic mass or mass number.

Examples of isotopes that can be incorporated into compounds ofaccording to Formula I include isotopes of hydrogen, carbon, nitrogen,oxygen, phosphorous, fluorine, chlorine, or iodine.

Illustrative of such isotopes are ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, O,¹⁷O, ¹⁸O, ³²P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I, and ¹²⁵I, respectively. Theseradiolabeled compounds can be used to measure the biodistribution,tissue concentration and the kinetics of transport and excretion frombiological tissues including a subject to which such a labeled compoundis administered. Labeled compounds are also used to determinetherapeutic effectiveness, the site or mode of action, and the bindingaffinity of a candidate therapeutic to a pharmacologically importanttarget. Certain radioactive-labeled compounds according to Formula I,therefore, are useful in drug and/or tissue distribution studies. Theradioactive isotopes tritium, i.e. ³H, and carbon-14, i.e. ¹⁴C, areparticularly useful for this purpose in view of their ease ofincorporation and ready means of detection.

Substitution with heavier isotopes such as deuterium, i.e. ²H, affordscertain therapeutic advantages resulting from the greater metabolicstability, for example, increased in vivo half-life of compoundscontaining deuterium. Substitution of hydrogen with deuterium may reducedose required for therapeutic effect, and hence may be preferred in adiscovery or clinical setting.

Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O and¹³N, provides labeled analogs of the inventive compounds that are usefulin Positron Emission Tomography (PET) studies, e.g., for examiningsubstrate receptor occupancy. Isotopically-labeled compounds accordingto Formula I, can generally be prepared by conventional techniques knownto those skilled in the art or by processes analogous to those describedin the Preparations and Examples section as set out below using anappropriate isotopic-labeling reagent.

Embodiments of the invention disclosed herein are also meant toencompass the in vivo metabolic products of compounds according toFormula I. Such products may result from, for example, the oxidation,reduction, hydrolysis, amidation, esterification, and like processesprimarily due to enzymatic activity upon administration of a compound ofthe invention. Accordingly, the invention includes compounds that areproduced as by-products of enzymatic or non-enzymatic activity on aninventive compound following the administration of such a compound to amammal for a period of time sufficient to yield a metabolic product.Metabolic products, particularly pharmaceutically active metabolites aretypically identified by administering a radiolabeled compound of theinvention in a detectable dose to a subject, such as rat, mouse, guineapig, monkey, or human, for a sufficient period of time during whichmetabolism occurs, and isolating the metabolic products from urine,blood or other biological samples that are obtained from the subjectreceiving the radiolabeled compound.

The invention also provides pharmaceutically acceptable salt forms ofFormula I compounds. Encompassed within the scope of the invention areboth acid and base addition salts that are formed by contacting apharmaceutically suitable acid or a pharmaceutically suitable base witha compound of the invention.

To this end, a “pharmaceutically acceptable acid addition salt” refersto those salts which retain the biological effectiveness and propertiesof the free bases, which are not biologically or otherwise undesirable,and which are formed with inorganic acids such as, but are not limitedto, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid and the like, and organic acids such as, but not limitedto, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid,ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid,4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid,capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid,citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonicacid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid,fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid,gluconic acid, glucuronic acid, glutamic acid, glutaric acid,2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuricacid, isobutyric acid, lactic acid, lactobionic acid, lauric acid,maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonicacid, mucic acid, naphthalene-1,5-disulfonic acid,naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid,oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid,propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid,4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid,tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroaceticacid, undecylenic acid, and the like.

Similarly, a “pharmaceutically acceptable base addition salt” refers tothose salts which retain the biological effectiveness and properties ofthe free acids, which are not biologically or otherwise undesirable.These salts are prepared by addition of an inorganic base or an organicbase to the free acid. Salts derived from inorganic bases include, butare not limited to, the sodium, potassium, lithium, ammonium, calcium,magnesium, iron, zinc, copper, manganese, aluminum salts and the like.Preferred inorganic salts are the ammonium, sodium, potassium, calcium,and magnesium salts. Salts derived from organic bases include, but arenot limited to, salts of primary, secondary, and tertiary amines,substituted amines including naturally occurring substituted amines,cyclic amines and basic ion exchange resins, such as ammonia,isopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, diethanolamine, ethanolamine, deanol,2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine,lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline,betaine, benethamine, benzathine, ethylenediamine, glucosamine,methylglucamine, theobromine, triethanolamine, tromethamine, purines,piperazine, piperidine. N-ethylpiperidine, polyamine resins and thelike. Particularly preferred organic bases are isopropylamine,diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, cholineand caffeine.

Often crystallizations produce a solvate of the compound of theinvention. As used herein, the term “solvate” refers to an aggregatethat comprises one or more molecules of a compound of the invention withone or more molecules of solvent. The solvent may be water, in whichcase the solvate may be a hydrate. Alternatively, the solvent may be anorganic solvent. Thus, the compounds of the present invention may existas a hydrate, including a monohydrate, dihydrate, hemihydrate,sesquihydrate, trihydrate, tetrahydrate and the like, as well as thecorresponding solvated forms. The compound of the invention may be truesolvates, while in other cases, the compound of the invention may merelyretain adventitious water or be a mixture of water plus someadventitious solvent.

A “stereoisomer” refers to a compound made up of the same atoms bondedby the same bonds but having different three-dimensional structures,which are not interchangeable. The present invention contemplatesvarious stereoisomers and mixtures thereof and includes “enantiomers”,which refers to two stereoisomers whose molecules are nonsuperimposeablemirror images of one another.

Compounds of the invention or their pharmaceutically acceptable saltsmay contain one or more asymmetric centers and may thus give rise toenantiomers, diastereomers, and other stereoisomeric forms that may bedefined, in terms of absolute stereochemistry, as (R)- or (S)- or, as(D)- or (L)-for amino acids. The present invention is meant to includeall such possible isomers, as well as their racemic and optically pureforms. Optically active (+) and (−). (R)- and (S)-, or (D)- and(L)-isomers may be prepared using chiral synthons or chiral reagents, orresolved using conventional techniques, for example, chromatography andfractional crystallization. Conventional techniques for thepreparation/isolation of individual enantiomers include chiral synthesisfrom a suitable optically pure precursor or resolution of the racemate(or the racemate of a salt or derivative) using, for example, chiralhigh pressure liquid chromatography (HPLC). When the compounds describedherein contain olefinic double bonds or other centers of geometricasymmetry, and unless specified otherwise, it is intended that thecompounds include both E and Z geometric isomers. Likewise, alltautomeric forms are also intended to be included.

The term “tautomer” refers to a proton shift from one atom of a moleculeto another atom of the same molecule. For example, when W¹ is oxo and A5is —NH, the present invention provides tautomers of a Formula I compoundas illustrated below:

The inventive compounds are synthesized using conventional syntheticmethods, and more specifically using the general methods noted below.Specific synthetic protocols for compounds in accordance with thepresent invention are described in the Examples.

Pharmaceutical Formulations

In one embodiment, a compounds according Formulae I are formulated aspharmaceutically acceptable compositions that contain a Formulae Icompound in an amount effective to treat a particular disease orcondition of interest upon administration of the pharmaceuticalcomposition to a mammal. Pharmaceutical compositions in accordance withthe present invention can comprise a Formulae I compound in combinationwith a pharmaceutically acceptable carrier, diluent or excipient.

In this regard, a “pharmaceutically acceptable carrier, diluent orexcipient” includes without limitation any adjuvant, carrier, excipient,glidant, sweetening agent, diluent, preservative, dye/colorant, flavorenhancer, surfactant, wetting agent, dispersing agent, suspending agent,stabilizer, isotonic agent, solvent, or emulsifier which has beenapproved by the United States Food and Drug Administration as beingacceptable for use in humans or domestic animals.

Further, a “mammal” includes humans and both domestic animals such aslaboratory animals and household pets (e.g., cats, dogs, swine, cattle,sheep, goats, horses, rabbits), and non-domestic animals such aswildlife and the like.

The pharmaceutical compositions of the invention can be prepared bycombining a compound of the invention with an appropriatepharmaceutically acceptable carrier, diluent or excipient, and may beformulated into preparations in solid, semi-solid, liquid or gaseousforms, such as tablets, capsules, powders, granules, ointments,solutions, suppositories, injections, inhalants, gels, microspheres, andaerosols. Typical routes of administering such pharmaceuticalcompositions include, without limitation, oral, topical, transdermal,inhalation, parenteral, sublingual, buccal, rectal, vaginal, andintranasal. The term parenteral as used herein includes subcutaneousinjections, intravenous, intramuscular, intrasternal injection orinfusion techniques. Pharmaceutical compositions of the invention areformulated so as to allow the active ingredients contained therein to bebioavailable upon administration of the composition to a patient.Compositions that will be administered to a subject or patient take theform of one or more dosage units, where for example, a tablet may be asingle dosage unit, and a container of a compound of the invention inaerosol form may hold a plurality of dosage units. Actual methods ofpreparing such dosage forms are known, or will be apparent, to thoseskilled in this art; for example, see Remington: The Science andPractice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy andScience, 2000). The composition to be administered will, in any event,contain a therapeutically effective amount of a compound of theinvention, or a pharmaceutically acceptable salt thereof, for treatmentof a disease or condition of interest in accordance with the teachingsof this invention.

A pharmaceutical composition of the invention may be in the form of asolid or liquid. In one aspect, the carrier(s) are particulate, so thatthe compositions are, for example, in tablet or powder form. Thecarrier(s) may be liquid, with the compositions being, for example, anoral syrup, injectable liquid or an aerosol, which is useful in, forexample, inhalatory administration. When intended for oraladministration, the pharmaceutical composition is preferably in eithersolid or liquid form, where semi-solid, semi-liquid, suspension and gelforms are included within the forms considered herein as either solid orliquid.

As a solid composition for oral administration the pharmaceuticalcomposition may be formulated into a powder, granule, compressed tablet,pill, capsule, chewing gum, wafer or the like form. Such a solidcomposition will typically contain one or more inert diluents or ediblecarriers. In addition, one or more of the following may be present:binders such as carboxymethylcellulose, ethyl cellulose,microcrystalline cellulose, gum tragacanth or gelatin; excipients suchas starch, lactose or dextrins, disintegrating agents such as alginicacid, sodium alginate. Primogel, corn starch and the like; lubricantssuch as magnesium stearate or Sterotex; glidants such as colloidalsilicon dioxide; sweetening agents such as sucrose or saccharin; aflavoring agent such as peppermint, methyl salicylate or orangeflavoring; and a coloring agent.

When the pharmaceutical composition is in the form of a capsule, forexample, a gelatin capsule, it may contain, in addition to materials ofthe above type, a liquid carrier such as polyethylene glycol or oil.

The pharmaceutical composition may be in the form of a liquid, forexample, an elixir, syrup, solution, emulsion or suspension. The liquidmay be for oral administration or for delivery by injection, as twoexamples. When intended for oral administration, preferred compositioncontain, in addition to the present compounds, one or more of asweetening agent, preservatives, dye/colorant and flavor enhancer. In acomposition intended to be administered by injection, one or more of asurfactant, preservative, wetting agent, dispersing agent, suspendingagent, buffer, stabilizer and isotonic agent may be included.

The liquid pharmaceutical compositions of the invention, whether they besolutions, suspensions or other like form, may include one or more ofthe following adjuvants: sterile diluents such as water for injection,saline solution, preferably physiological saline, Ringer's solution,isotonic sodium chloride, fixed oils such as synthetic mono ordiglycerides which may serve as the solvent or suspending medium,polyethylene glycols, glycerin, propylene glycol or other solvents:antibacterial agents such as benzyl alcohol or methyl paraben;antioxidants such as ascorbic acid or sodium bisulfite; chelating agentssuch as ethylenediaminetetraacetic acid; buffers such as acetates,citrates or phosphates and agents for the adjustment of tonicity such assodium chloride or dextrose. The parenteral preparation can be enclosedin ampoules, disposable syringes or multiple dose vials made of glass orplastic. Physiological saline is a preferred adjuvant. An injectablepharmaceutical composition is preferably sterile.

A liquid pharmaceutical composition of the invention intended for eitherparenteral or oral administration should contain an amount of a compoundof the invention such that a suitable dosage will be obtained.

The pharmaceutical composition of the invention may be intended fortopical administration, in which case the carrier may suitably comprisea solution, emulsion, ointment or gel base. The base, for example, maycomprise one or more of the following: petrolatum, lanolin, polyethyleneglycols, bee wax, mineral oil, diluents such as water and alcohol, andemulsifiers and stabilizers. Thickening agents may be present in apharmaceutical composition for topical administration. If intended fortransdermal administration, the composition may include a transdermalpatch or iontophoresis device.

The pharmaceutical composition of the invention may be intended forrectal administration, in the form, for example, of a suppository, whichwill melt in the rectum and release the drug. The composition for rectaladministration may contain an oleaginous base as a suitablenonirritating excipient. Such bases include, without limitation,lanolin, cocoa butter and polyethylene glycol.

The pharmaceutical composition of the invention may include variousmaterials, which modify the physical form of a solid or liquid dosageunit. For example, the composition may include materials that form acoating shell around the active ingredients. The materials that form thecoating shell are typically inert, and may be selected from, forexample, sugar, shellac, and other enteric coating agents.Alternatively, the active ingredients may be encased in a gelatincapsule.

The pharmaceutical composition of the invention in solid or liquid formmay include an agent that binds to the compound of the invention andthereby assists in the delivery of the compound. Suitable agents thatmay act in this capacity include a monoclonal or polyclonal antibody, aprotein or a liposome.

The pharmaceutical composition of the invention may consist of dosageunits that can be administered as an aerosol. The term aerosol is usedto denote a variety of systems ranging from those of colloidal nature tosystems consisting of pressurized packages. Delivery may be by aliquefied or compressed gas or by a suitable pump system that dispensesthe active ingredients.

Aerosols of compounds of the invention may be delivered in single phase,bi-phasic, or tri-phasic systems in order to deliver the activeingredient(s). Delivery of the aerosol includes the necessary container,activators, valves, subcontainers, and the like, which together may forma kit. One skilled in the art, without undue experimentation maydetermine preferred aerosols.

The pharmaceutical compositions of the invention may be prepared by anymethodology well known in the pharmaceutical art. For example, apharmaceutical composition intended to be administered by injection canbe prepared by combining a compound of the invention with sterile,distilled water so as to form a solution. A surfactant may be added tofacilitate the formation of a homogeneous solution or suspension.Surfactants are compounds that non-covalently interact with the compoundof the invention so as to facilitate dissolution or homogeneoussuspension of the compound in the aqueous delivery system.

In certain embodiments a pharmaceutical composition comprising acompound of Formula I is administered to a mammal in an amountsufficient to inhibit Mnk activity upon administration, and preferablywith acceptable toxicity to the same. Mnk activity of Formula Icompounds can be determined by one skilled in the art, for example, asdescribed in the Examples below. Appropriate concentrations and dosagescan be readily determined by one skilled in the art.

Therapeutic Use

The compounds of the invention, or their pharmaceutically acceptablesalts, are administered in a therapeutically effective amount, whichwill vary depending upon a variety of factors including the activity ofthe specific compound employed; the metabolic stability and length ofaction of the compound; the age, body weight, general health, sex, anddiet of the patient; the mode and time of administration; the rate ofexcretion; the drug combination; the severity of the particular disorderor condition; and the subject undergoing therapy.

“Effective amount” or “therapeutically effective amount” refers to thatamount of a compound of the invention which, when administered to amammal, preferably a human, is sufficient to effect treatment, asdefined below, of a Mnk related condition or disease in the mammal,preferably a human. The amount of a compound of the invention whichconstitutes a “therapeutically effective amount” will vary depending onthe compound, the condition and its severity, the manner ofadministration, and the age of the mammal to be treated, but can bedetermined routinely by one of ordinary skill in the art having regardto his own knowledge and to this disclosure.

Compounds of the invention or pharmaceutically acceptable salt thereofmay also be administered simultaneously with, prior to, or afteradministration of one or more other therapeutic agents. Such combinationtherapy includes administration of a single pharmaceutical dosageformulation which contains a compound of the invention and one or moreadditional active agents, as well as administration of the compound ofthe invention and each active agent in its own separate pharmaceuticaldosage formulation. For example, a compound of the invention and theother active agent can be administered to the patient together in asingle oral dosage composition such as a tablet or capsule, or eachagent administered in separate oral dosage formulations. Where separatedosage formulations are used, the compounds of the invention and one ormore additional active agents can be administered at essentially thesame time, i.e., concurrently, or at separately staggered times, i.e.,sequentially; combination therapy is understood to include all theseregimens.

In certain embodiments the disclosed compounds are useful for inhibitingthe activity of Mnk and/or can be useful in analyzing Mnk signalingactivity in model systems and/or for preventing, treating, orameliorating a symptom associated with a disease, disorder, orpathological condition involving Mnk, preferably one afflicting humans.A compound which inhibits the activity of Mnk will be useful inpreventing, treating, ameliorating, or reducing the symptoms orprogression of diseases of uncontrolled cell growth, proliferationand/or survival, inappropriate cellular immune responses, orinappropriate cellular inflammatory responses or diseases which areaccompanied with uncontrolled cell growth, proliferation and/orsurvival, inappropriate cellular immune responses, or inappropriatecellular inflammatory responses, particularly in which the uncontrolledcell growth, proliferation and/or survival, inappropriate cellularimmune responses, or inappropriate cellular inflammatory responses ismediated by Mnk, such as, for example, haematological tumors, solidtumors, and/or metastases thereof, including leukaemias andmyelodysplastic syndrome. Waldenstrom macroglobulinemia, and malignantlymphomas, for example, B-cell lymphoma. T-cell lymphoma, hairy celllymphoma. Hodgkins lymphoma, non-Hodgins lymphoma, and Burkettslymphoma, head and neck tumors including brain tumors and brainmetastases, tumors of the thorax including non-small cell and small celllung tumors, gastrointestinal tumors, endocrine tumors, mammary andother gynecological tumors, urological tumors including renal, bladderand prostate tumors, skin tumors, and sarcomas, and/or metastasesthereof.

Furthermore, the inventive compounds and their pharmaceuticalcompositions are candidate therapeutics for the prophylaxis and/ortherapy of cytokine related diseases, such as inflammatory diseases,allergies, or other conditions associated with proinflammatorycytokines. Exemplary inflammatory diseases include without limitation,chronic or acute inflammation, inflammation of the joints such aschronic inflammatory arthritis, rheumatoid arthritis, psoriaticarthritis, osteoarthritis, juvenile rheumatoid arthritis, Reiter'ssyndrome, rheumatoid traumatic arthritis, rubella arthritis, acutesynovitis and gouty arthritis; inflammatory skin diseases such assunburn, psoriasis, erythrodermic psoriasis, pustular psoriasis, eczema,dermatitis, acute or chronic graft formation, atopic dermatitis, contactdermatitis, urticaria and scleroderma; inflammation of thegastrointestinal tract such as inflammatory bowel disease, Crohn'sdisease and related conditions, ulcerative colitis, colitis, anddiverticulitis; nephritis, urethritis, salpingitis, oophoritis,endomyometritis, spondylitis, systemic lupus erythematosus and relateddisorders, multiple sclerosis, asthma, meningitis, myelitis,encephalomyelitis, encephalitis, phlebitis, thrombophlebitis,respiratory diseases such as asthma, bronchitis, chronic obstructivepulmonary disease (COPD), inflammatory lung disease and adultrespiratory distress syndrome, and allergic rhinitis; endocarditis,ostcomyelitis, rheumatic fever, rheumatic pericarditis, rheumaticendocarditis, rheumatic myocarditis, rheumatic mitral valve disease,rheumatic aortic valve disease, prostatitis, prostatocystitis,spondoarthropathies ankylosing spondylitis, synovitis, tenosynovotis,myositis, pharyngitis, polymyalgia rheumatica, shoulder tendonitis orbursitis, gout, pseudo gout, vasculitides, inflammatory diseases of thethyroid selected from granulomatous thyroiditis, lymphocyticthyroiditis, invasive fibrous thyroiditis, acute thyroiditis;Hashimoto's thyroiditis, Kawasaki's disease, Raynaud's phenomenon,Sjogren's syndrome, neuroinflammatory disease, sepsis, conjunctivitis,keratitis, iridocyclitis, optic neuritis, otitis, lymphoadenitis,nasopaharingitis, sinusitis, pharyngitis, tonsillitis, laryngitis,epiglottitis, bronchitis, pneumonitis, stomatitis, gingivitis,oesophagitis, gastritis, peritonitis, hepatitis, cholelithiasis,cholecystitis, glomerulonephritis, goodpasture's disease, crescenticglomerulonephritis, pancreatitis, endomyometritis, myometritis,metritis, cervicitis, endocervicitis, exocervicitis, parametritis,tuberculosis, vaginitis, vulvitis, silicosis, sarcoidosis,pneumoconiosis, pyresis, inflammatory polyarthropathies, psoriatricarthropathies, intestinal fibrosis, bronchiectasis and enteropathicarthropathies.

Although inflammation is the unifying pathogenic process of thesediseases, current therapies only treat the symptoms of the disease andnot the underlying cause of inflammation. The compositions of thepresent invention are useful for the treatment and/or prophylaxis ofinflammatory diseases and related complications and disorders.

Accordingly, certain embodiments are directed to a method for treating aMnk dependent condition in a mammal in need thereof, the methodcomprising administering an effective amount of a pharmaceuticalcomposition as described above (i.e., a pharmaceutical compositioncomprising any one or more compounds of Formula I) to a mammal.

As described above deregulation of protein synthesis is a common eventin human cancers. A key regulator of translational control is eIF4Ewhose activity is a key determinant of tumorigenicity. Becauseactivation of eIF4E involves phosphorylation of a key serine (Ser209)specifically by MAP kinase interacting kinases (Mnk), inhibitors of Mnkare suitable candidate therapeutics for treating cell proliferativedisorders such as cancer. A wide variety of cancers, including solidtumors, lymphomas and leukemias, are amenable to the compositions andmethods disclosed herein. Types of cancer that may be treated include,but are not limited to: adenocarcinoma of the breast, prostate andcolon; all forms of bronchogenic carcinoma of the lung; myeloid;melanoma; hepatoma; neuroblastoma; papilloma; apudoma; choristoma;branchioma; malignant carcinoid syndrome: carcinoid heart disease: andcarcinoma (e.g., Walker, basal cell, basosquamous, Brown-Pearce, ductal,Ehrlich tumor, Krebs 2, merkel cell, mucinous, non-small cell lung, oatcell, papillary, scirrhous, bronchiolar, bronchogenic, squamous cell,and transitional cell). Additional types of cancers that may be treatedinclude: histiocytic disorders; acute and chronic leukemia, both myeloidand lymphoid/lymphoblastic, including hairy cell leukemia; histiocytosismalignant; Hodgkins disease; immunoproliferative small; Hodgkinslymphoma; B-cell and T-cell non-Hodgkins lymphoma, including diffuselarge B-cell and Burkett's lymphoma; plasmacytoma:reticuloendotheliosis; melanoma; multiple myeloma; chondroblastoma;chondroma; chondrosarcoma; fibroma; fibrosarcoma; myelofibrosis; giantcell tumors; histiocytoma; lipoma: liposarcoma: mesothelioma; myxoma;myxosarcoma; osteoma; osteosarcoma; chordoma; craniopharyngioma;dysgerminoma; hamartoma; mesenchymoma; mesonephroma: myosarcoma;ameloblastoma; cementoma; odontoma; teratoma; thymoma; trophoblastictumor.

Other cancers that can be treated using the inventive compounds includewithout limitation adenoma; cholangioma; cholesteatoma; cyclindroma;cystadenocarcinoma; cystadenoma; granulosa cell tumor; gynandroblastoma;hepatoma; hidradenoma; islet cell tumor; Leydig cell tumor; papilloma;sertoli cell tumor, theca cell tumor; leimyoma; leiomyosarcoma;myoblastoma; myomma; myosarcoma; rhabdomyoma; rhabdomyosarcoma;ependymoma; ganglioneuroma; glioma; medulloblastoma; meningioma;neurilemmoma; neuroblastoma; neuroepithelioma; neurofibroma; neuroma;paraganglioma; paraganglioma nonchromaffin.

In one embodiment the inventive compounds are candidate therapeuticagents for the treatment of cancers such as angiokeratoma; angiolymphoidhyperplasia with eosinophilia; angioma sclerosing; angiomatosis;glomangioma; hemangioendothelioma; hemangioma; hemangiopericytoma;hemangiosarcoma; lymphangioma; lymphangiomyoma; lymphangiosarcoma;pinealoma; carcinosarcoma; chondrosarcoma; cystosarcoma phyllodes;fibrosarcoma; hemangiosarcoma; leiomyosarcoma; leukosarcoma:liposarcoma: lymphangiosarcoma; myosarcoma; myxosarcoma; ovariancarcinoma; rhabdomyosarcoma; sarcoma; neoplasms; nerofibromatosis; andcervical dysplasia.

In a particular embodiment the present disclosure provides methods fortreating colon cancer, colorectal cancer, gastric cancer, thyroidcancer, lung cancer, leukemia, pancreatic cancer, melanoma, multiplemelanoma, brain cancer, primary and secondary CNS cancer, includingmalignant glioma and glioblastoma, renal cancer, prostate cancer,including castration-resistant prostate cancer, ovarian cancer, orbreast cancer, including triple negative, HER2 positive, and hormonereceptor positive breast cancers. According to such a method, atherapeutically effective amount of at least one compound according toFormula I or a stereoisomer, tautomer or pharmaceutically acceptablesalt thereof can be administered to a subject who has been diagnosedwith a cell proliferative disease, such as a cancer. Alternatively, apharmaceutical composition comprising at least one compound according toFormula I or a stereoisomer, tautomer or pharmaceutically acceptablesalt thereof can be administered to a subject who has been diagnosedwith cancer.

In certain embodiments the compounds in accordance with the inventionare administered to a subject with cancer in conjunction with otherconventional cancer therapies such as radiation treatment or surgery.Radiation therapy is well-known in the art and includes X-ray therapies,such as gamma-irradiation, and radiopharmaceutical therapies.

In certain embodiments the inventive Mnk inhibitor compounds are usedwith at least one anti-cancer agent. Anti-cancer agents includechemotherapeutic drugs. A chemotherapeutic agent includes, but is notlimited to, an inhibitor of chromatin function, a topoisomeraseinhibitor, a microtubule inhibiting drug, a DNA damaging agent, anantimetabolite (such as folate antagonists, pyrimidine analogs, purineanalogs, and sugar-modified analogs), a DNA synthesis inhibitor, a DNAinteractive agent (such as an intercalating agent), and a DNA repairinhibitor.

Illustrative chemotherapeutic agents include, without limitation, thefollowing groups: anti-metabolites/anti-cancer agents, such aspyrimidine analogs (5-fluorouracil, floxuridine, capecitabine,gemcitabine and cytarabine) and purine analogs, folate antagonists andrelated inhibitors (mercaptopurine, thioguanine, pentostatin and2-chlorodeoxyadenosine (cladribine)); antiproliferative/antimitoticagents including natural products such as vinca alkaloids (vinblastine,vincristine, and vinorelbine), microtubule disruptors such as taxane(paclitaxel, docetaxel), vincristin, vinblastin, nocodazole, epothilonesand navelbine, epidipodophyllotoxins (etoposide, teniposide), DNAdamaging agents (actinomycin, amsacrine, anthracyclines, bleomycin,busulfan, camptothecin, carboplatin, chlorambucil, cisplatin,cyclophosphamide, Cytoxan, dactinomycin, daunorubicin, doxorubicin,epirubicin, hexamethylmelamineoxaliplatin, iphosphamide, melphalan,merchlorehtamine, mitomycin, mitoxantrone, nitrosourea, plicamycin,procarbazine, taxol, taxotere, temozolamide, teniposide,triethylenethiophosphoramide and etoposide (VP 16)); antibiotics such asdactinomycin (actinomycin D), daunorubicin, doxorubicin (adriamycin),idarubicin, anthracyclines, mitoxantrone, bleomycins, plicamycin(mithramycin) and mitomycin; enzymes (L-asparaginase which systemicallymetabolizes L-asparagine and deprives cells which do not have thecapacity to synthesize their own asparagine); antiplatelet agents;antiproliferative/antimitotic alkylating agents such as nitrogenmustards (mechlorethamine, cyclophosphamide and analogs, melphalan,chlorambucil), ethylenimines and methylmelamines (hexamethylmelamine andthiotepa), alkyl sulfonates—busulfan, nitrosoureas (carmustine (BCNU)and analogs, streptozocin), trazenes-dacarbazinine (DTIC);antiproliferative/antimitotic antimetabolites such as folic acid analogs(methotrexate); platinum coordination complexes (cisplatin,carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide;hormones, hormone analogs (estrogen, tamoxifen, goserelin, bicalutamide,nilutamide) and aromatase inhibitors (letrozole, anastrozole);anticoagulants (heparin, synthetic heparin salts and other inhibitors ofthrombin): fibrinolytic agents (such as tissue plasminogen activator,streptokinase and urokinase), aspirin, dipyridamole, ticlopidine,clopidogrel, abciximab; antimigratory agents: antisecretory agents(breveldin); immunosuppressives (cyclosporine, tacrolimus (FK-506),sirolimus (rapamycin), azathioprine, mycophenolate mofetil);anti-angiogenic compounds (TNP470, genistein) and growth factorinhibitors (vascular endothelial growth factor (VEGF) inhibitors,fibroblast growth factor (FGF) inhibitors); angiotensin receptorblocker; nitric oxide donors; anti-sense oligonucleotides; antibodies(trastuzumab, rituximab); chimeric antigen receptors; cell cycleinhibitors and differentiation inducers (tretinoin); mTOR inhibitors,topoisomerase inhibitors (doxorubicin (adriamycin), amsacrine,camptothecin, daunorubicin, dactinomycin, eniposide, epirubicin,etoposide, idarubicin, irinotecan (CPT-11) and mitoxantrone, topotecan,irinotecan), corticosteroids (cortisone, dexamethasone, hydrocortisone,methylpednisolone, prednisone, and prenisolone); growth factor signaltransduction kinase inhibitors; mitochondrial dysfunction inducers,toxins such as Cholera toxin, ricin. Pseudomonas exotoxin. Bordetellapertussis adenylate cyclase toxin, or diphtheria toxin, and caspaseactivators; and chromatin disruptors.

In certain embodiments an Mnk inhibitor in accordance with the presentinvention is used simultaneously, in the same formulation or in separateformulations, or sequentially with an additional agent(s) as part of acombination therapy regimen.

Mnk inhibitors according to Formula I including their correspondingsalts and pharmaceutically acceptable compositions of Formula Icompounds are also effective as therapeutic agents for treating orpreventing cytokine mediated disorders, such as inflammation in apatient, preferably in a human. In one embodiment, a compound orcomposition in accordance with the invention is particularly useful fortreating or preventing a disease selected from chronic or acuteinflammation, chronic inflammatory arthritis, rheumatoid arthritis,psoriasis, COPD, inflammatory bowel disease, septic shock, Crohn'sdisease, ulcerative colitis, multiple sclerosis and asthma.

The inventive compounds their corresponding salts and pharmaceuticallyacceptable compositions are candidate therapeutics for treating brainrelated disorders which include without limitation autism, FragileX-syndrome. Parkinson's disease and Alzheimer's disease.

Treatment is effected by administering to a subject in need of treatmenta Formula I compound, its pharmaceutically acceptable salt form, or apharmaceutically acceptable composition of a Formula I compound or itssalt.

The invention also supports the use of the inventive compounds or apharmaceutically acceptable formulation of the inventive compound as aninhibitor of Mnk activity. Such inhibition is achieved by contacting acell expressing Mnk with a compound or a pharmaceutically acceptableformulation, to lower or inhibit Mnk activity, to provide therapeuticefficacy for a Mnk dependent condition in a mammal in need thereof.

Therapeutically effective dosages of a compound according to Formula Ior a composition of a Formula I compound will generally range from about1 to 2000 mg/day, from about 10 to about 1000 mg/day, from about 10 toabout 500 mg/day, from about 10 to about 250 mg/day, from about 10 toabout 100 mg/day, or from about 10 to about 50 mg/day. Thetherapeutically effective dosages may be administered in one or multipledoses. It will be appreciated, however, that specific doses of thecompounds of the invention for any particular patient will depend on avariety of factors such as age, sex, body weight, general healthcondition, diet, individual response of the patient to be treated, timeof administration, severity of the disease to be treated, the activityof particular compound applied, dosage form, mode of application andconcomitant medication. The therapeutically effective amount for a givensituation will readily be determined by routine experimentation and iswithin the skills and judgment of the ordinary clinician or physician.In any case the compound or composition will be administered at dosagesand in a manner which allows a therapeutically effective amount to bedelivered based upon patient's unique condition.

General Synthetic Methods Method 1:

The formation of I is accomplished by reacting compound II (P¹ is anoptional protecting group) with compound III (X is a leaving group, suchas halogen, —OTf, —OTs or -OMs, and P² is an optional protecting group)under the Buchwald-Hartwig conditions (such as palladium catalyst,ligand, base, solvent and heat), followed by de-protection and/orfurther functional group manipulation if necessary.

Alternatively, coupling of compound II (P¹ is an optional protectinggroup) and compound III (X is a leaving group, such as halogen, —OTf,—OTs or -OMs, and P² is an optional protecting group) is alsoaccomplished under the copper-mediated Ullmann type conditions (such ascopper(I) iodide, base, solvent, heat), followed by de-protection and/orfurther functional group manipulation if necessary.

Method 2:

The formation of I is also accomplished by reacting compound II (P¹ isan optional protecting group) with compound IV (R is a hydrogen or alkylgroup and P² is an optional protecting group) under the Chan-Lamconditions (such as copper(II) acetate, oxygen, base, solvent, heat),followed by de-protection and/or further functional group manipulationif necessary.

Method 3:

Additionally, the formation of I is also accomplished by reactingcompound II (P¹ is an optional protecting group) with compound V (P² isan optional protecting group) under the typical nucleophilic aromaticsubstitution conditions (such as solvent, heat), followed byde-protection and/or further functional group manipulation if necessary.

Method 4:

The formation of intermediate VII is accomplished by exposing compoundVI (when P is an optional protecting group) to an alkyl halide underbasic conditions (such as sodium hydride in tetrahydrofuran), followedby de-protection and/or further functional group manipulation ifnecessary.

Method 5:

The formation of IX is accomplished by exposing VIII (when P is anoptional protecting group) to the Wittig olefination conditions (such asPh₃═P—CR^(9a)R^(9b), solvent and heat), followed by de-protection and/orfurther functional group manipulation if necessary.

Synthesis of Formula I Compounds

The following examples are provided for purpose of illustration and notlimitation.

Example 1 Synthesis of5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 1F)

Synthesis of 2-(4-methoxybenzyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (3)

Procedure A: To a solution of5-bromo-2-(4-methoxybenzyl)isoindolin-1-one (1, 0.5 g, 1.5 mmol) indioxane (10 mL) was added 7H-pyrrolo[2,3-d]pyrimidine (2, 0.27 g, 2.25mmol) and potassium tert-butoxide (0.0.51 g, 4.52 mmol) followed by theaddition of XantPhos (0.087 g, 0.15 mmol). The reaction mixture wasdegassed with argon for 15 min. Tris(dibenzylideneacetone)dipalladium(0)(0.14 g, 0.15 mmol) was then added and the reaction mixture was heatedat 90° C. and maintained at that temperature for 12 h.

Following heating, the reaction mixture was cooled and concentratedunder reduced pressure. The concentrated reaction mixture was extractedin ethyl acetate. The organic layer was separated, dried over sodiumsulphate, filtered and concentrated under reduced pressure. The residueobtained was purified by silica gel (100-200 mesh) column chromatographyusing 5% methanol in dichloromethane as eluent so as to afford2-(4-methoxybenzyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(3). Yield: 0.21 g, 38%; MS (ESI) m/z 371[M+1]⁺.

Synthesis of 5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd.No. F)

Procedure B: A solution of2-(4-methoxybenzyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(3, 0.21 g, 0.55 mmol) in trifluoroacetic acid (10 mL) was heated at 95°C. for 12 h. Following heating the reaction mixture was concentratedunder reduced pressure and neutralized with saturated solution of sodiumbicarbonate. The residue thus obtained was filtered, washed with waterand then with hexane and diethyl ether. The solid was dried to afford5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 1F).Yield: 0.08 g, 58%; MS (ESI) m/z 251[M+1]⁺; ¹H NMR (400 MHz. DMSO-d₆) δ9.16 (s, 1H), 8.92 (s, 1H), 8.68 (s, 1H), 8.21-8.11 (m, 2H), 8.04 (d,J=8.2 Hz, 1H), 7.85 (d, J=8.2 Hz, 1H), 6.94 (d, J=3.8 Hz, 1H), 4.49 (s,2H).

Example 2 Synthesis of3-methyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd. No.2F)

Synthesis of 5-bromo-2-(4-methoxybenzyl)-3-methylisoindolin-1-one (2)

To a stirring solution of 5-bromo-2-(4-methoxybenzyl)isoindolin-1-one(1.1 g, 3 mmol) in tetrahydrofuran (10 mL) at 0° C., was added sodiumhydride (0.14 g, 3.62 mmol) in small portions. The resultant reactionmixture was allowed warm to room temperature and stirred at roomtemperature for an additional 10 min. Iodomethane (0.64 g, 4.51 mmol)was then added and the reaction mixture was refluxed for 1 h, thenquenched with water and extracted with ethyl acetate. The organic layerwas separated, dried over sodium sulphate and concentrated under reducedpressure. The residue thus obtained was purified by silica gel columnchromatography using 20% ethyl acetate in hexane as eluent to afford5-bromo-2-(4-methoxybenzyl)-3-methylisoindolin-1-one (2). Yield: 0.3 g,29%; MS (ESI) m/z 347[M+1]⁺.

Synthesis of2-(4-methoxybenzyl)-3-methyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(4)

The synthesis of intermediate 4 was carried out according to the generalprotocol described in Procedure A. Yield: 0.1 g, 45%; MS (ESI) nm/z385[M+1]⁺.

Synthesis of3-methyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd. No.2F)

The synthesis of compound 2F was carried out according to the generalprotocol described in Procedure B. Yield: 0.05 g, 74%; MS (ESI) nm/z265[M+1]⁺; ¹H NMR (400 MHz. DMSO-d₆) δ 9.16 (s, 1H), 8.91 (s, 1H), 8.77(s, 1H), 8.15 (d, J=3.8 Hz, 2H), 8.07 (dd, J=8.1, 2.0 Hz, 1H), 7.82 (d,J=8.2 Hz, 1H), 6.94 (d, J=3.8 Hz, 1H), 4.73 (q, J=6.7 Hz, 1H), 1.44 (d,J=6.7 Hz, 3H).

Example 3 Synthesis of3-methyl-5-(5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)isoindolin-1-one (Cpd.No. 3)

Synthesis of3-methyl-5-(SH-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)isoindolin-1-one (Cpd.No. 3)

To a solution of3-methyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (1, 0.07 g,0.26 mmol) in methanol (5 mL), was added 10% palladium on carbon (70mg). The reaction mixture was allowed to stir at room temperature underan atmosphere of hydrogen for 48 h, then filtered through celite andwashed with methanol. The filtrate was concentrated under reducedpressure and the residue was purified by flash chromatography to afford3-methyl-5-(5H-pyrrolo[2,3-d]pyrimidin-7(6H)-yl)isoindolin-1-one (Cpd.No. 3). Yield: 0.017 g, 25%; MS (ESI) n/z 267[M+1]⁺: ¹H NMR (400 MHz.DMSO-d₆) δ 8.57 (s, 1H), 8.49 (s, 1H), 8.24 (d, J=1.4 Hz, 1H), 8.09-8.01(m, 2H), 7.65 (d, J=8.2 Hz, 1H), 4.63 (q. J=6.7 Hz, 1H), 4.21-4.17 (m,2H), 3.20 (t, J=8.7 Hz, 2H), 1.38 (d, J=6.6 Hz, 3H).

Example 4 Synthesis of5-(1H-pyrazolo[4,3-c]pyridin-1-yl)isoindolin-1-one (Cpd. No. 4)

Synthesis of2-(4-methoxybenzyl)-5-(1H-pyrazolo[4,3-c]pyridin-1-yl)isoindolin-1-one(3)

To a solution of 5-bromo-2-(4-methoxybenzyl)isoindolin-1-one (1, 0.25 g,2.09 mmol) in dimethyl sulfoxide (5 mL), was added1H-pyrazolo[4,3-c]pyridine (2, 0.84 g, 2.51 mmol), and copper(I) iodide(0.08 g, 0.42 mmol) followed by the addition of cesium carbonate (1.35g, 4.18 mmol). The reaction mixture was heated at 120° C. for 20 h thendiluted with ethyl acetate and filtered through celite. The filtrate wasevaporated under reduced pressure and the residue was purified by silicagel column chromatography using 10% methanol in dichloromethane aseluent to afford2-(4-methoxybenzyl)-5-(1H-pyrazolo[4,3-c]pyridin-1-yl)isoindolin-1-one(3). Yield: 0.5 g, 64%; MS (ESI) m/z 371[M+1]⁺.

Synthesis of 5-(1H-pyrazolo[4,3-c]pyridin-1-yl)isoindolin-1-one (Cpd.No. 4)

The synthesis of compound 4 was carried out according to the generalprotocol described in Procedure B. Yield: 0.05 g, 15%; MS (ESI) m/z251[M+1]⁺; ¹H NMR (400 MHz. DMSO-d₆) δ 9.26 (s, 1H), 8.68 (d, J=3.6 Hz,2H), 8.54 (d, J=6.1 Hz, 1H), 8.07-8.02 (m, 1H), 8.00-7.84 (m, 3H), 4.50(s, 2H).

Example 5 Synthesis of2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Cpd. No. 5)

Synthesis of6-(4-methoxybenzyl)-2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrrolo([3,4-b]pyridin-5-one(3)

The synthesis of intermediate 3 was carried out according to the generalprotocol described in Procedure A. Yield: 0.15 g, 23%; MS (ESI) m/z371.05[M+1]⁺.

Synthesis of 2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one (Cpd. No. 5)

The synthesis of compound 5 was carried out according to the generalprotocol described in Procedure B. Yield: 0.028 g, 27%; MS (ESI) nm/z251[M+1]⁺; ¹H NMR (400 MHz. DMSO-d₆) δ 8.95 (s, 1H), 8.78 (s, 1H),8.47-8.37 (m, 2H), 8.26 (dd, J=6.1, 2.4 Hz, 2H), 7.93 (d, J=8.4 Hz, 1H),6.98 (d, J=3.5 Hz, 1H), 4.55 (s, 2H).

Example 6 Synthesis of 6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoquinoline(Cpd. No. 6)

Synthesis of 6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoquinoline (Cpd. No.6)

The synthesis of compound 6 was carried out according to the generalprotocol described in Procedure A. Yield: 0.042 g, 13%; MS (ESI) nm/z247[M+1]⁺; ¹H NMR (400 MHz. DMSO-(d₆) 9.39 (s, 1H), 9.19 (s, 1H), 8.96(s, 1H), 8.60-8.50 (m, 2H), 8.39-8.22 (m, 3H), 7.93 (d, J=5.8 Hz, 1H),6.99 (d, J=3.8 Hz, 1H).

Example 7 Synthesis of 5-(4-methyl-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 7)

Synthesis of 2-(4-methoxybenzyl)-5-(4-methyl-7H-pyrrolo[2, 3-d]pyrimidin-7-yl) isoindolin-1-one (3)

Procedure C: A solution of 4-methyl-7H-pyrrolo[2,3-d]pyrimidine (1, 0.4g, 3.0 mmol), 5-bromo-2-(4-methoxybenzyl) isoindolin-1-one (2, 1.0 g,4.5 mmol) and potassium phosphate (1.91 g, 9.0 mmol) in 1, 4-dioxane (15ml) was degassed with nitrogen for 10 min. Copper (I) iodide (0.28 g,1.5 mmol) and trans-1, 2-diaminocyclohexane (0.17 g, 1.5 mmol) wereadded and the reaction was refluxed at 90° C. for 16 h. Progress of thereaction was monitored by TLC. After completion, solvent was removedunder reduced pressure. The reaction mixture was diluted with water andextracted twice with ethyl acetate. The organic layer was separated,dried over sodium sulphate, filtered and concentrated under reducedpressure to afford 2-(4-methoxybenzyl)-5-(4-methyl-7H-pyrrolo[2, 3-d]pyrimidin-7-yl) isoindolin-1-one (3) as a yellow solid. Yield: 0.55 g,47%; MS (ESI) m/z 385.12[M+1]⁺.

Synthesis of 5-(4-methyl-7H-pyrrolo[2, 3-d] pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 7)

The synthesis of compound 7 was carried out according to the generalprotocol described in Procedure B. White solid; Yield: 0.058 g, 21%; MS(ESI) m/z 265.07[M+1]⁺; ¹H NMR (400 MHz. DMSO-d₆) δ 8.76 (s, 1H), 8.66(s, 1H), 8.17 (s, 1H), 8.07 (d, J=3.6 Hz, 1H), 8.03 (d, J=8.4 Hz, 1H),7.84 (d, J=8.0 Hz, 1H), 6.99 (d, J=4.0 Hz, 1H), 4.48 (s, 2H), 2.73 (s,3H).

Example 8 Synthesis of5-(5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(Cpd. No. 8)

Synthesis of5-bromo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(2)

To a solution of 5-bromo-7H-pyrrolo[2,3-d]pyrimidine (1, 0.9 g, 4.54mmol) in tetrahydrofuran (10 mL) at 0° C., was added sodium hydride(0.27 g, 6.81 mmol, 60% in hexane). The reaction mixture was allowed tostir at 0° C. for 20 min. Chloromethyl 2-trimethylsilylethyl ether (0.9g, 5.44 mmol) was then added and the reaction mixture was stirred at 0°C. for an additional 30 min., and then quenched with water. The solventwas removed under reduced pressure and the residue was purified bysilica gel column chromatography using 5% methanol in dichloromethane toafford5-bromo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(2). Yield: 1.3 g, 86%; MS (ESI) m/z 328[M+1]⁺.

Synthesis of5-(1-methyl-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(4)

Procedure D: To a solution of5-bromo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(2, 0.5 g, 1.51 mmol) in 1,4-dioxane and water (15 mL, 4:1), was added1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (3,0.41 g, 1.97 mmol) and sodium carbonate (0.48 g, 4.53 mmol). Thereaction mixture was degassed with argon for 15 min., and then[1,1-bis(diphenylphosphino)ferrocene] dichloropalladium (0.18 g, 0.22mmol) was added and the reaction mixture heated at 90° C. for 16 h.After heating, the reaction mixture was diluted with ethyl acetate andfiltered through celite. The filtrate was evaporated under reducedpressure and the residue was purified by silica gel columnchromatography using 5% methanol in dichloromethane as eluent to afford5-(1-methyl-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(4). Yield: 0.7 g, 70%; MS (ESI) m/z 330[M+1]⁺.

Synthesis of 5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine(5)

To a solution of5-(1-methyl-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine (4, 0.7 g, 2.12 mmol) indichloromethane (10 mL), was added cold trifluoroacetic acid (10 mL) at0° C. The reaction mixture was warmed and allowed to stir at roomtemperature for 16 h, following which it was concentrated under reducedpressure. The residue thus obtained was diluted with acetonitrile andaqueous ammonia and stirred at room temperature for 1 h. After stirringthe reaction mixture was concentrated under reduced pressure and theresidue purified by neutral silica gel (100-200 mesh) columnchromatography using 5% methanol in dichloromethane as eluent to afford5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine (5). Yield:0.42 g, 98%.

Synthesis of2-(4-methoxybenzyl)-5-(5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(7)

The synthesis of intermediate 7 was carried out according to the generalprotocol described in Procedure A. Yield: 0.3 g, 33%; MS (ESI) m/z451[M+1]⁺.

Synthesis of5-(5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(Cpd. No. 8)

Procedure E: To a solution of2-(4-methoxybenzyl)-5-(5-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(7, 0.05 g, 0.11 mmol) in toluene (2 mL), was added triflic acid (0.2mL, 0.44 mmol) at room temperature and the reaction mixture was heatedin a microwave at 140° C. for 15 min. Following heating the reactionmixture was concentrated under reduced pressure and neutralized using asaturated solution of sodium bicarbonate. The precipitated solid wasfiltered, washed with dichloromethane, followed by pentane, methanol anddiethyl ether. The washed residue was dried to afford5-(5-(l-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(Cpd. No. 8). Yield: 0.03 g, 14%; MS (ESI) n/z 331[M+1]⁺; ¹H NMR (400MHz. DMSO-d₆) δ 9.51 (s, 1H), 9.03 (s, 1H), 8.70 (s, 1H), 8.49 (s, 1H),8.39 (s, 1H), 8.22 (s, 1H), 8.12-8.02 (m, 2H), 7.89 (d, J=8.2 Hz, 1H),4.50 (s, 2H), 3.93 (s, 3H).

Example 9 Synthesis of5-(6,7-dihydropyrido[2,3-d]pyrimidin-8(5H)-yl)-3-methylisoindolin-1-one(Cpd. No. 9)

Synthesis of5-(6,7-dihydropyrido[2,3-d]pyrimidin-8(5H)-yl)-2-(4-methoxybenzyl)-3-methylisoindolin-1-one(3)

The synthesis of intermediate 3 was carried out according to the generalprotocol described in Procedure A. Yield: 0.18 g, 51%; MS (ESI) m/z401[M+1]⁺.

Synthesis of5-(6,7-dihydropyrido[2,3-d]pyrimidin-8(5H)-yl)-3-methylisoindolin-1-one(Cpd. No. 9)

The synthesis of compound 9 was carried out according to the generalprotocol described in Procedure E. Yield: 0.03 g, 43%; MS (ESI) nm/z281[M+1]⁺; ¹H NMR (400 MHz, Methanol-d₄) δ 8.27 (s, 1H), 8.06 (s, 1H),7.83 (d, J=8.1 Hz, 1H), 7.59 (d, J=1.8 Hz, 1H), 7.49 (dd, J=8.1, 1.8 Hz,1H), 4.88 (t, J=2.5 Hz, 1H), 3.95-3.91 (m, 2H), 2.91 (t, J=6.0 Hz, 2H),2.16 (m, 2H), 1.49 (d, J=6.7 Hz, 3H), 1.31 (d, J=14.1 Hz, 1H).

Example 10 Synthesis of4-(1-oxoisoindolin-5-yl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one(Cpd. No. 10)

Synthesis of tert-butyl4-chloro-1-oxo-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate (2)

To a solution of 4-chloro-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (1,150 mg, 0.89 mmol) in tetrahydrofuran (1 mL) was added4-(dimethylamino)pyridine (11 mg, 0.09 mmol) and di-tert-butyldicarbonate (194 mg, 0.89 mmol). The reaction was stirred at roomtemperature for 30 min. The resulting mixture was concentrated andpurified via column chromatography (silica, ethyl acetate/hexanes=0-20%)to afford tert-butyl4-chloro-1-oxo-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate (2)as an off-white solid. Yield: 186 mg, 78%.

Synthesis of tert-butyl4-(2-(tert-butoxycarbonyl)-1-oxoisoindolin-5-yl)-1-oxo-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate(4)

The synthesis of intermediate 4 was carried out according to the generalprotocol described in Procedure D. Yield: 58 mg, 26%.

Synthesis of 4-(1-oxoisoindolin-5-yl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (Cpd. No. 10)

To a suspension of tert-butyl4-(2-(tert-butoxycarbonyl)-1-oxoisoindolin-5-yl)-1-oxo-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate(4, 39 mg, 0.08 mmol) in methanol (4 mL) was added a 4 M solution ofhydrogen chloride in dioxane (4 mL, 16 mmol). The reaction was stirredat room temperature for 90 min, and then at 50° C. for 30 min. Uponcooling, the mixture was concentrated and triturated with ethyl acetateto afford4-(1-oxoisoindolin-5-yl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one(Cpd. No. 10). Yield: 15 mg, 59%. ¹H NMR (300 MHz, DMSO-d₆) δ 9.20 (s,1H), 8.88 (d, J=4.8 Hz, 1H), 8.70 (s, 1H), 8.14 (d, J=0.6 Hz, 1H), 8.06(dd. J=1.5, 7.5 Hz, 1H), 7.81 (d, J=8.1 Hz, 1H), 7.73 (d, J=4.5 Hz, 1H),4.79 (s, 2H), 4.47 (s, 2H).

Example 11 Synthesis of6-(1H-pyrrolo[3,2-c]pyridin-1-yl)-1H-pyrrolo[3,4-c]pyridin-3(2H)-one(Cpd. No. 11)

Synthesis of6-chloro-2-(4-methoxybenzyl)-1H-pyrrolo/3,4pyridin-3(21-1)-one

To a stirring solution of 2,2,6,6-tetramethylpiperidine (10 g, 71.2mmol) in tetrahydrofuran (10 mL) was added n-butyl lithium (44 mL, 71.2mmol) at −78° C. To this solution were added2-((4-methoxybenzyl)amino)acetonitrile (1.4 g 17.8 mmol) intetrahydrofuran (15 mL) followed by the addition of(6-chloropyridin-3-yl)(piperidin-1-yl)methanone (2 3.76 g, 21.38 mmol)in tetrahydrofuran (15 mL) at −78° C. Stirring of the reaction mixturewas continued at −78° C. for 7 h. The progress of reaction was monitoredby TLC. After complete consumption of starting material, reaction wasquenched with ammonium chloride solution and extracted with ethylacetate. The organic layer was separated, washed with brine, dried oversodium sulfate and concentrated under reduced pressure to a cruderesidue, which was purified by column chromatography to afford6-chloro-2-(4-methoxybenzyl)-1H-pyrrolo [3,4-c]pyridin-3(2H)-one (3).Yield: 1.2 g, 23%; MS (ESI) m/z 289[M+1]⁺.

Synthesis of2-(4-methoxybenzyl)-6-(1H-pyrrolo[3,2-c]pyridin-I-yl)-1H-pyrrolo[3,4-c]pyridin-3(2H)-one(5)

The synthesis of intermediate 5 was carried according to the generalprotocol described in Procedure A. Yield: 0.31 g, 34%; MS (ESI) nm/z 372[M+1]⁺.

Synthesis of6-(1H-pyrrolo[3,2-c]pyridin-1-yl)-1H-pyrrolo[3,4-c]pyridin-3(2H)-one(Cpd. No. 11)

The synthesis of compound II was carried out according to the generalprotocol described in Procedure E. Yield: 0.045 g, 26%; MS (ESI) nm/z251[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.96 (s, 1H), 8.89 (s, 1H), 8.78(s, 1H), 8.40 (s, 2H), 8.23 (d, J=3.6 Hz, 1H), 8.09 (s, 1H), 6.98 (d,J=3.6 Hz, 1H), 4.56 (s, 2H).

Example 12 Synthesis of 5-(4-amino-7H-pyrrolo [2, 3-d] pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 12)

Synthesis of N-(4-methoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (2)

A mixture of 4-chloro-7H-pyrrolo [2, 3-d] pyrimidine (1, 2.0 g, 13.07mmol), 4-methoxybenzylamine (3.6 g, 26.14 mmol) and potassium carbonate(5.42 g, 39.21 mmol) in 1,4-dioxane (20 ml) was refluxed at 100° C. for16 h. Progress of the reaction was monitored by TLC. After completion,solvent was removed under reduced pressure, the reaction mixture wasdiluted with water and extracted with ethyl acetate twice. The organiclayer was again washed with brine, separated, dried over sodium sulphateand concentrated under reduced pressure. The residue was finally washedwith pentane to affordN-(4-methoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (2) as an offwhite solid. Yield: 0.82 g, 25%; MS (ESI) m/z 254.99[M+1]⁺; H NMR (400MHz. DMSO-d₆) δ 11.49 (s, 1H), 8.09 (s, 1H), 7.85 (t, J=6.0 Hz, 1H),7.27 (d, J=8.8 Hz, 2H), 7.06 (s, 1H), 6.87 (d, J=8.8 Hz, 2H), 6.57 (s,1H), 4.63 (d, J=6.0 Hz, 2H), 3.71 (s, 3H).

Synthesis of 2-(4-methoxybenzyl)-5-(4-((4-methoxybenzyl)amino)-7H-pyrrolo [2,3-d]pyrimidin-7-yl)isoindolin-1-one (4)

The synthesis of intermediate 4 was carried out according to the generalprotocol described in Procedure A. Off-white solid; Yield: 0.63 g, 40%;MS (ESI) m/z 506.35[M+1]⁺.

Synthesis of 5-(4-amino-7H-pyrrolo [2, 3-d] pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 12)

The synthesis of compound 12 was carried out according to the generalprotocol described in Procedure B. Off-white solid; Yield: 0.07 g, 24%;MS (ESI) m/z 266.07[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.61 (s, 1H),8.15 (s, 2H), 7.99 (s, 1H), 7.79 (s, 1H), 7.66 (s, 1H), 7.21 (s, 2H),6.82 (s, 1H), 4.45 (s, 2H).

Example 13 Synthesis of7-(1,3-dihydrobenzo[c]thiophen-5-yl)-7H-pyrrolo[2,3-d]pyrimidine (Cpd.No. 13)

Synthesis of7-(1,3-dihydrobenzo[c]thiophen-5-yl)-7H-pyrrolo[2,3-d]pyrimidine (Cpd.No. 13)

To a solution of 5-bromo-1,3-dihydrobenzo[c]thiophene (1, 0.18 g, 0.81mmol) in dimethylformamide (3.5 mL), were added 7H-pyrrolo[2,3-d]pyrimidine (2.0.14 g, 1.22 mmol) and sodium tert-butoxide (0.12g, 1.2 mmol). The reaction mixture was degassed with argon for 20 min.Trans-N,N′-dimethyl cyclohexane 1,2 diamine (0.064 g, 0.4 mmol) andcopper(I) iodide (0.030 g, 0.16 mmol) were then added and the reactionmixture was heated at 100° C. for 16 h. After heating the reactionmixture was filtered through celite and the filtrate was concentratedunder reduced pressure. The residue thus obtained was purified by silicagel column chromatography using 2% methanol in dichloromethane as eluentto afford7-(1,3-dihydrobenzo[c]thiophen-5-yl)-7H-pyrrolo[2,3-d]pyrimidine (Cpd.No. 13). Yield: 0.029 g, 12%; MS (ESI) m/z 254[M+1]⁺; ¹H NMR (400 MHz,DMSO-d₆) δ 9.13 (s, 1H), 8.87 (s, 1H), 8.03 (d, J=3.7 Hz, 1H), 7.84 (d,J=2.0 Hz, 1H), 7.74 (dd. J=8.3, 2.1 Hz, 1H), 7.50 (d, J=8.2 Hz, 1H),6.88 (d, J=3.7 Hz, 1H), 4.34-4.26 (m, 4H).

Example 14 Synthesis of2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-5-one(Cpd. No. 14)

Synthesis of ethyl 4-formyl-2-(methylthio)pyrimidine-5-carboxylate (2)

To a stirring solution of ethyl4-methyl-2-(methylthio)pyrimidine-5-carboxylate (1, 2.6 g, 12.24 mmol)in 1,4-dioxane (65 mL), was added selenium dioxide (2.71 g, 24.49 mmol).The reaction mixture was heated at 100° C. for 24 h, and progress of thereaction was monitored by TLC. After completion of the reaction, themixture was cooled and filtered. The filtrate was concentrated underreduced pressure to afford ethyl4-formyl-2-(methylthio)pyrimidine-5-carboxylate (2). Yield: 2.5 g, 90%;MS (ESI) m/z 227[M+1]⁺.

Synthesis of 6-(4-methoxybenzyl)-2-(methylthio)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-5-one (3)

To a solution of ethyl 4-formyl-2-(methylthio)pyrimidine-5-carboxylate(2, 2.5 g, 11.04 mmol) in methanol (25 mL) and dichloromethane (25 mL),was added dropwise a solution of 4-methoxybenzylamine (1.51 g, 11.04mmol). The reaction mixture was stirred at room temperature for 30 min.Sodium cyanoborohydride (1.73 g, 27.6 mmol) was then added and thereaction mixture was stirred at room temperature for an additional 24 h.The progress of the reaction was monitored by TLC. After completeconsumption of starting material, the reaction mixture was concentratedunder reduced pressure and the residue was diluted with water and thecompound was extracted with dichloromethane. The organic layer waswashed with brine, dried over anhydrous sodium sulfate and concentratedunder reduced pressure. The residue was purified by columnchromatography to6-(4-methoxybenzyl)-2-(methylthio)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-5-one(3). Yield: 1.6 g, 48%; MS (ESI) m/z 302[M+1]⁺.

Synthesis of 6-(4-methoxybenzyl)-2-(methylsulfonyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-5-one (4)

To a solution of6-(4-methoxybenzyl)-2-(methylthio)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-5-one(3, 1.6 g, 5.30 mmol) in dichloromethane (160 mL) at 0° C., was added insmall portions m-chloroperbenzoic acid (2.74 g, 15.92 mmol) over aperiod of 30 min. The reaction mixture was stirred at room temperaturefor 1 h. The progress of the reaction was monitored by TLC. Aftercomplete consumption of starting material the reaction mixture wasquenched with saturated solution of sodium bicarbonate and extractedwith dichloromethane. The organic layer was washed with brine, driedover anhydrous sodium sulfate and concentrated under reduced pressure.The resultant residue was purified by repeated washing with ether andpentane to give6-(4-methoxybenzyl)-2-(methylsulfonyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-5-one(4). Yield: 0.45 g, 26%; MS (ESI) m/z 334[M+1]⁺.

Synthesis of 6-(4-methoxybenzyl)-2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-51H-pyrrolo[3,4-d]pyrimidin-5-one (6)

To a solution of6-(4-methoxybenzyl)-2-(methylsulfonyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-5-one(4, 0.2 g, 0.60 mmol) in acetonitrile (5 mL), was added at 0° C.1H-pyrrolo[3,2-c]pyridine (5, 0.049 g, 0.42 mmol) and the reactionmixture was stirred at room temperature for 1 h. The progress of thereaction was monitored by TLC. After complete consumption of startingmaterial, the reaction mixture was concentrated under reduced pressureand the residue was purified by column chromatography to afford6-(4-methoxybenzyl)-2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-5-one(6). Yield: 0.1 g, 22%; MS (ESI) m/z 372[M+1]⁺.

Synthesis of2-(1H-pyrrolo[3,2-c]pyridin-1-yl)-6,7-dihydro-SH-pyrrolo[3,4-d]pyrimidin-5-one,formic acid salt (Cpd. No. 14)

The synthesis of compound 14 was carried out according to the generalprotocol described in Procedure B. Yield: 0.009 g, 13%; MS (ESI) m/z252.10[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.18 (s, 1H), 8.97 (s, 1H),8.90 (s, 1H), 8.65 (d, J=5.8 Hz, 1H), 8.51-8.36 (m, 1H), 7.00 (d, J=3.7Hz, 1H), 4.61 (s, 2H), 2.67-2.51 (m, 1H).

Example 15 Synthesis of 5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)indolin-2-one(Cpd. No. 15)

Synthesis of ethyl2-(2-nitro-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)acetate (3)

The synthesis of intermediate 3 was carried out according to the generalprotocol described in Procedure A. Yield: 0.4 g, 35%.

Synthesis of ethyl2-(2-amino-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)acetate (4)

To a solution of ethyl2-(2-nitro-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)acetate (3, 0.35 g,1.07 mmol) in ethanol (25 mL), was added 10% palladium on carbon (150mg) and the reaction mixture was allowed to stir at room temperatureunder an atmosphere of hydrogen for 4 h. The reaction mixture wasfiltered through a celite bed and the filtrate was concentrated underreduced pressure to afford ethyl2-(2-amino-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)acetate (4). Yield:0.38 g, crude.

Synthesis of 5-(7H-pyrrolo[2, 3-d]pyrimidin-7-yl)indolin-2-one (Cpd. No.15)

A solution of ethyl2-(2-amino-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)acetate (4, (0.35g, 1.18 mmol) in ethanol (20 mL) was allowed to reflux for 16 h. Thereaction mixture was concentrated under reduced pressure and the residuewas purified by neutral silica gel column chromatography using 5%methanol in dichloromethane as eluent to afford compound5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)indolin-2-one (Cpd. No. 15). Thecompound was lyophilised to remove the trapped methanol and trituratedwith diethyl ether to remove the non polar impurity. Yield: 0.058 g,17%. MS (ESI) m/z 251[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 10.56 (s, 1H),9.11 (s, 1H), 8.83 (s, 1H), 7.92 (d, J=3.9 Hz, 1H), 7.68 (s, 1H), 7.58(d, J=8.3 Hz, 1H), 6.97 (d, J=8.3 Hz, 1H), 6.89-6.80 (m, 1H), 3.60 (s,2H).

Example 16 Synthesis of 5-(9H-purin-9-yl) isoindolin-1-one (Cpd. No. 16)

Synthesis of2-(4-methoxybenzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-1-one(2)

A mixture of 5-bromo-2-(4-methoxybenzyl) isoindolin-1-one (1, 1.0 g,3.02 mmol), bis(pinacolato) diboron (0.84 g, 3.32 mmol) and potassiumacetate (0.74 g, 7.55 mmol) in 1,4-dioxane (10 ml) was degassed withargon at room temperature for 15 min. Then[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.22 g,0.30 mmol) was added under nitrogen atmosphere and the reaction waspurged for another 10 min. The reaction was allowed to reflux at 100° C.for 18 h. Progress of the reaction was monitored by TLC. Aftercompletion, the reaction mass was filtered through celite and the celitebed was washed with ethyl acetate. The combined organic layer was driedover sodium sulphate and concentrated under reduced pressure to afford2-(4-methoxybenzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-1-one(2) as a black solid. Yield: 1.2 g, crude: MS (ESI) m/z 380.27[M+1⁺].

Synthesis of mixture of 2-(4-methoxybenzyl)-5-(9H-purin-9-yl)isoindolin-1-one (4) and 2-(4-methoxybenzyl)-5-(7H-purin-7-yl)isoindolin-1-one (4a)

A stirred solution of2-(4-methoxybenzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-1-one(2, 1.26 g, 3.34 mmol), 9H-purine (3, 0.2 g, 1.67 mmol) andtetramethylethylenediamine (0.39 g, 3.34 mmol) in methanol (30 ml) andwater (5 ml) was degassed with oxygen for 10 min. Copper (II) acetate(0.31 g, 1.67 mmol) was then added and the reaction was stirred at roomtemperature for 16 h under an atmosphere of oxygen. Progress of thereaction was monitored by TLC. After completion, solvent was removedunder reduced pressure. The reaction mixture was diluted with water andextracted twice with ethyl acetate. The organic layer was separated,dried over sodium sulphate and concentrated under reduced pressure. Theresidue was purified via column chromatography using 5% methanol indichloromethane to afford a mixture of2-(4-methoxybenzyl)-5-(9H-purin-9-yl) isoindolin-1-one (4) and2-(4-methoxybenzyl)-5-(7H-purin-7-yl) isoindolin-1-one (4a) as a brownsolid. Yield: 0.46 gm, 36%; MS (ESI) m/z 372.05 and 372.09[M+1]⁺.

Synthesis of 5-(9H-purin-9-yl) isoindolin-1-one (Cpd. No. 16)

The synthesis of compound 16 was carried out according to the generalprotocol described in Procedure B. White solid; Yield: 0.075 g, 52%; MS(ESI) m/z 252.03[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.33 (s, 1H), 9.14(s, 1H), 9.06 (s, 1H), 8.77 (s, 1H), 8.23 (s, 1H), 8.08 (d, J=8.4 Hz,1H), 7.91 (d, J=8.0 Hz, 1H), 4.51 (s, 2H).

Example 17 Synthesis of 7-chloro-3-methyl-5-(7H-pyrrolo [2, 3-d]pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 17)

Synthesis of methyl 4-bromo-2-(bromomethyl)-6-chlorobenzoate (2)

To a solution of methyl 4-bromo-2-chloro-6-methylbenzoate (1, 2.0 g, 7.6mmol) in carbon tetrachloride (20 ml) were added N-bromosuccinimide (1.6g, 9.1 mmol) and 2,2′-azobis(2-methylpropionitrile) (0.25 g, 1.52 mmol).The reaction mixture was stirred at 80° C. for 12 h and progress of thereaction was monitored by TLC. After completion, solvent was dilutedwith dichloromethane and the organic layer was washed with water andbrine. Following separation, the organic layer was dried using sodiumsulphate and concentrated under reduced pressure to afford methyl4-bromo-2-(bromomethyl)-6-chlorobenzoate (2) as a yellow sticky liquid.Yield: 3.9 g, crude.

Synthesis of 5-bromo-7-chloro-2-(4-methoxybenzyl) isoindolin-1-one (3)

To a solution of methyl 4-bromo-2-(bromomethyl)-6-chlorobenzoate (2, 3.0g, 8.8 mmol) in N,N-dimethylformamide (25 ml) were added4-methoxybenzylamine (1.8 g, 13.0 mmol) and triethylamine (2.67 g, 26.4mmol). The reaction was stirred at room temperature for 12 h andprogress of reaction was monitored by TLC. After completion, thereaction mass was quenched with water and the desired compound wasextracted from the crude reaction mass with ethyl acetate twice. Thecombined organic layers were washed with cold water and cold brine.Following separation, the organic layer was dried using sodium sulphateand concentrated under reduced pressure. The residue was purified viacolumn chromatography using 10% ethyl acetate in hexane to get5-bromo-7-chloro-2-(4-methoxybenzyl) isoindolin-1-one (3) as a yellowsticky liquid. Yield: 1.6 g, 50%; MS (ESI) m/z 365.97[M+1]⁺; ¹H NMR (400MHz, DMSO-d₆) δ 7.77 (d, J=4.4 Hz, 2H), 7.22 (d, J=8.8 Hz, 2H), 6.91 (d,J=8.4 Hz, 2H), 4.61 (s, 2H), 4.29 (s, 2H), 3.73 (s, 3H).

Synthesis of mixture ofS-bromo-7-chloro-2-(4-methoxybenzyl)-3-methylisoindolin-1-one (4) and5-bromo-7-chloro-2-(4-methoxybenzyl)-3,3-dimethylisoindolin-1-one (4′)

To a solution of 5-bromo-7-chloro-2-(4-methoxybenzyl) isoindolin-1-one(3, 1.5 g, 4.1 mmol) in dry tetrahydrofuran (20 mL) at 0° C. was addedsolid sodium bis(trimethylsilyl)amide (0.89 g, 4.92 mmol). The reactionmass was stirred at 0° C. for 15 min. Iodomethane (5.8 g, 41.0 mmol) wasthen added and the reaction mixture was stirred at 0° C. for anadditional 3 h. Progress of the reaction was monitored by TLC. Afterconsumption of starting material, the reaction mixture was quenched withwater and extracted twice with ethyl acetate. The organic layer waswashed with brine solution, dried over anhydrous sodium acetate,filtered and concentrated under reduced pressure to get a mixture of5-bromo-7-chloro-2-(4-methoxybenzyl)-3-methylisoindolin-1-one (4) and5-bromo-7-chloro-2-(4-methoxybenzyl)-3,3-dimethylisoindolin-1-one (4′)as a yellow sticky liquid. Yield: 0.85 g, crude; MS (ESI) m/z380.02[M+1] and 394.04[M+1]⁺.

Synthesis of mixture of7-chloro-2-(4-methoxybenzyl)-3-methyl-(7H-pyrrolo [2,3-d]pyrimidin-7-yl) isoindolin-1-one (6) and7-chloro-2-(4-methoxybenzyl)-3, 3-dimethyl-5-(7H-pyrrolo [2, 3-d]pyrimidin-7-yl) isoindolin-1-one (6′)

The synthesis of a mixture of 6 and 6′ was carried out according to thegeneral protocol described in Procedure C. Brown solid; Yield: 0.45 g,crude; MS (ESI) m/z 419.27[M+1]⁺ and 433.28[M+1]⁺.

Synthesis of 7-chloro-3-methyl-5-(7H-pyrrolo [2, 3-d] pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 17)

The synthesis of compound 17 was carried out according to the generalprotocol described in Procedure B. Yield: 0.057 g, 40%; MS (ESI) m/z299.02[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.17 (s, 1H), 8.96 (s, 1H),8.87 (s, 1H), 8.22 (d, J=4.0 Hz, 2H), 8.18 (s, 1H), 6.96 (d, J=3.6 Hz,1H), 4.70 (m, 1H), 1.43 (d, J=6.4 Hz, 3H).

Example 18 Synthesis of 7-chloro-3, 3-dimethyl-5-(7H-pyrrolo [2, 3-d]pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 18)

Synthesis of 7-chloro-3, 3-dimethyl-5-(7H-pyrrolo [2, 3-d]pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 18)

The synthesis of compound 18 was carried out as described above inExample 17. Yield: 0.025 g, 17%; MS (ESI) m/z 313.02[M+1]⁺. ¹H NMR (400MHz. DMSO-d₆) δ 9.17 (s, 1H), 8.96 (s, 1H), 8.87 (s, 1H), 8.26 (s, 1H),8.23 (d, J=3.6 Hz, 1H), 8.17 (s, 1H), 6.96 (d, J=4.0 Hz, 1H), 1.51 (s,6H).

Example 19 Synthesis of5-(5-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(Cpd. No. 19)

Synthesis of 5-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(2)

To a solution of 5-(7H-pyrrolo [2,3-d] pyrimidin-7-yl) isoindolin-1-one(1, 1.3 g, 5.17 mmol) in N,N-dimethylformamide (15 ml) was addedN-bromosuccinimide (1.04 g, 5.69 mmol) and the reaction mixture wasstirred at room temperature for 1 h. Progress of the reaction wasmonitored by TLC. After completion, the reaction mass was quenched withwater and extracted twice with ethyl acetate. The organic layer was thenseparated, dried over sodium sulphate and concentrated under reducedpressure to afford5-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (2) as anoff white solid. Yield: 0.85 g, 50%; MS (ESI) m/z 329.01[M+1]⁺; ¹H NMR(400 MHz. DMSO-d₆) δ 9.09 (s, 1H), 9.00 (s, 1H), 8.70 (s, 1H), 8.45 (s,1H), 8.15 (s, 1H), 8.00 (d, J=8.0 Hz, 1H), 7.85 (d, J=8.0 Hz, 1H), 4.48(s, 2H).

Synthesis of 5-(5-(pyridin-4-yl)-7H-pyrrolo [2, 3-d] pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 19)

The synthesis of compound 19 was carried out according to the generalprotocol described in Procedure D. Off white solid; Yield: 0.018 g, 12%;MS (ESI) nm/z 328.06[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.65 (s, 1H),9.02 (s, 1H), 8.88 (s, 1H), 8.72 (s, 1H), 8.66 (d, J=5.6 Hz, 2H), 8.25(s, 1H), 8.11 (d, J=8.4 Hz, 1H), 7.97 (d, J=5.6 Hz, 2H), 7.90 (d, J=8.4Hz, 1H), 4.52 (s, 2H).

Example 20 Synthesis of methyl((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbamate(Cpd. No. 20)

Synthesis of ethyl2-(6-bromo-2-(4-methoxybenzyl)-3-oxoisoindolin-1-yl)acetate (3)

To a solution of 5-bromo-2-(4-methoxybenzyl)isoindolin-1-one (1, 1 g,3.01 mmol) in tetrahydrofuran (25 mL) at −78° C. was added dropwise asolution of sodium bis(trimethylsilyl)amide (552 mg, 3.01 mmol) intetrahydrofuran (15 mL). The reaction was stirred at −78° C. for 15 min,followed by the dropwise addition of ethyl 2-bromoacetate (2, 503 mg,3.01 mmol) in tetrahydrofuran (10 mL). The reaction was stirred at −78°C. for an additional 20 min before it was warmed to room temperaturegradually. The warmed reaction mixture is then poured into a halfsaturated ammonium chloride solution and the aqueous solution isextracted with ethyl acetate. The ethyl acetate layers were combined,washed with brine, dried over magnesium sulfate, filtered andconcentrated under reduced pressure. The crude product was purified viacolumn chromatography (silica, ethyl acetate/hexanes=0-10%) to affordethyl 2-(6-bromo-2-(4-methoxybenzyl)-3-oxoisoindolin-1-yl)acetate (3).Yield: 572 mg, 45%; MS (ESI) m/z 418.3[M+1]⁺.

Synthesis of ethyl2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)acetate(5)

A mixture of ethyl2-(6-bromo-2-(4-methoxybenzyl)-3-oxoisoindolin-1-yl)acetate (3, 439 mg,1.05 mmol), 7H-pyrrolo[2,3-d]pyrimidine (4, 125 mg, 1.05 mmol),tris(dibenzylideneacetone)dipalladium(00 (97 mg, 0.10 mmol), XantPhos(61 mg, 0.10 mmol), and cesium carbonate (752 mg, 2.31 mmol) in1,4-dioxane (25 mL) was purged with argon for 5 min. The reaction wasstirred at 110° C. for 16 h. Upon cooling, the reaction mixture wasdiluted with ethyl acetate, washed with half saturated aqueous sodiumbicarbonate solution, and then with brine. The organic layer was driedover magnesium sulfate, filtered and concentrated. The crude product waspurified via column chromatography (silica, methanol/dichloromethanegradient from 0-5% to afford ethyl2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)acetate(5). Yield: 333 mg, 70%; MS (ESI) m/z 457.4[M+1]⁺.

Synthesis of2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)aceticacid (6)

To a solution of ethyl2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)acetate(4, 333 mg, 0.73 mmol) in methanol (10 mL) and water (5 mL) was addedlithium hydroxide (52 mg, 2.19 mmol). The reaction was stirred at roomtemperature for 2 h and then acidified to pH ˜5 with 1.25 M hydrogenchloride in ethanol. The resulting mixture was concentrated to afford2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)aceticacid (6). Yield: 314 mg, 100%; MS (ESI) m/z 429.2[M+1]⁺.

Synthesis of methyl((2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-S-yl)methyl)carbamate(7)

To a suspension of2-(2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)aceticacid (6, 312 mg, 0.73 mmol) in tetrahydrofuran (10 mL) at 0° C. wasadded triethylamine (0.41 mL, 2.92 mmol), and ethyl chloroformate (119mg, 1.09 mmol). The reaction was stirred at 0° C. for 1 h. A solution ofsodium azide (95 mg, 1.46 mmol) in water (2 mL) was then added to thereaction mixture. After stirring at 0° C. for 5 min the reaction mixturewas warmed to room temperature and allowed to stir for an additional 2h. The resulting mixture was poured into water and extracted with ethylacetate. The separated organic layer was washed with half saturatedaqueous sodium bicarbonate solution (20 mL), and brine. It was thendried over magnesium sulfate, filtered and concentrated. The residuethus obtained was dissolved in dichloromethane (15 mL) and refluxed for30 min. The reaction was cooled to room temperature followed by theaddition of methanol (1 mL). The reaction was stirred at reflux for anadditional 1 h. The resulting mixture was concentrated and purified viacolumn chromatography (silica, methanol/dichloromethane=0-5%) to affordmethyl((2-(4-methoxybenzyl)-3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbamate(7). Yield: 181 mg, 54%; MS (ESI) m/z 458.5[M+1]⁺.

Synthesis of methyl((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbamate(Cpd. No. 20)

The synthesis of compound 20 was carried out according to the generalprotocol described in Procedure B. White solid; Yield: 34 mg, 42%; MS(ESI) m/z 338.3[M+1]⁺; ¹H NMR (300 MHz, CD₃OD) δ 9.42 (s, 1H), 9.15 (s,1H), 8.31 (d, J=3.9 Hz, 1H), 8.13 (s, 1H), 8.07-7.98 (m, 2H), 7.25 (d,J=3.6 Hz, 1H), 4.92 (s, 1H), 3.72-3.50 (m, 5H).

Example 21 Synthesis of3-(aminomethyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-onehydrochloride (Cpd. No. 21)

Synthesis of tert-butyl((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbamate(2)

To a solution of methyl((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbamate(1, 22 mg, 0.07 mmol) in acetonitrile (5 mL) was addediodotrimethylsilane (0.02 mL, 0.13 mmol). The reaction was stirred atroom temperature for 2 h. The resulting mixture was concentrated andre-dissolved in dichloromethane (5 mL), followed by the addition ofdi-tert-butyl dicarbonate (29 mg, 0.13 mmol). After stirring at roomtemperature for 4 h, the mixture was concentrated and purified viacolumn chromatography (silica, methanol/dichloromethane=0-10%) to affordtert-butyl((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbamate(2). Yield: 18 mg, 73%; MS (ESI) m/z 380.2[M+1]⁺.

Synthesis of3-(aminomethyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-onehydrochloride (Cpd. No. 21)

To a solution of tert-butyl((3-oxo-6-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-yl)methyl)carbamate(2, 18 mg, 0.05 mmol) in methanol (6 mL) was added 4 M hydrogen chloridein dioxane (0.01 mL, 0.05 mmol). The reaction was stirred at roomtemperature for 2 h and the resulting mixture was concentrated andtriturated with methanol and ether to afford3-(aminomethyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-onehydrochloride (Cpd. No. 21) as a white solid. Yield: 8 mg, 50%; MS (ESI)m/z 280.4[M+1]⁺. ¹H NMR (300 MHz, CD₃OD) δ 9.50 (s, 1H), 9.22 (s, 1H),8.39 (d, J=3.6 Hz, 1H), 8.27-8.26 (m, 1H), 8.17 (dd, J=7.8, 1.5 Hz, 1H),8.08 (d, J=8.4 Hz, 1H), 7.33 (d, J=3.9 Hz, 1H), 5.18 (t, J=4.2 Hz, 1H),3.70-3.51 (m, 2H).

Example 22 Synthesis of3,7-dimethyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd.No. 22)

Synthesis of 5-bromo-2-(4-methoxybenzyl)-3, 7-dimethylisoindolin-1-one(2)

To a solution of 5-bromo-2-(4-methoxybenzyl)-7-methylisoindolin-1-one(1, 2.2 g, 6.3 mmol) in tetrahydrofuran (20 mL) at 0° C. was added solidsodium bis(trimethylsilyl)amide (1.39 g, 7.6 mmol). The reaction wasstirred at 0° C. for 15 min. Iodomethane (1.17 g, 8.2 mmol) was thenadded and reaction mixture stirred at 0° C. for an additional 4 h.Progress of the reaction was monitored by TLC. After consumption ofstarting material, the reaction mixture was quenched with water andextracted with ethyl acetate. After separation of the organic layer fromthe aqueous layer, the former was washed with brine, dried overanhydrous sodium sulfate, filtered and concentrated under reducedpressure to afford5-bromo-2-(4-methoxybenzyl)-3,7-dimethylisoindolin-1-one (2) as a whitesolid. Yield: 1.0 g, 44%; MS (ESI) m/z 360.16[M+1]⁺.

Synthesis of 2-(4-methoxybenzyl)-3,7-dimethyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (4)

The synthesis of intermediate 4 was carried out according to the generalprotocol described in Procedure C. Off white solid: Yield: 0.41 g, 93%.MS (ESI) m/z 399.24[M+1]⁺.

Synthesis of 3,7-dimethyl-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one (Cpd. No.22)

The synthesis of compound 22 was carried out according to the generalprotocol described in Procedure B. Off white solid; Yield: 0.14 g, 57%.MS (ESI) m/z 279.09[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.150 (s, 1H),8.916 (s, 1H), 8.639 (s, 1H), 8.110-8.119 (d, J=3.6 Hz, 1H), 7.937 (s,1H), 7.791 (s, 1H), 6.915-6.924 (d, J=3.8 Hz, 1H), 4.67 (m, 1H), 2.685(s, 3H), 1.397-1.414 (d, J=6.8 Hz, 3H).

Example 23 Synthesis of 7-fluoro-5-(7H-pyrrolo [2, 3-d] pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 23)

Synthesis of methyl 4-bromo-2-(bromomethyl)-6-fluorobenzoate (2)

To a solution of methyl 4-bromo-2-fluoro-6-methylbenzoate (1, 1.8 g,7.31 mmol) in carbon tetrachloride (100 ml) were addedN-bromosuccinimide (1.56 g, 8.78 mmol) and2,2′-azobis(2-methylpropionitrile) (0.24 g, 1.46 mmol). The reaction wasrefluxed at 80° C. for 18 h and the progress was monitored by TLC. Aftercompletion, solvent was removed under reduced pressure to afford methyl4-bromo-2-(bromomethyl)-6-fluorobenzoate (2) as a brown solid. Yield:3.9 g, crude; ¹H NMR (400 MHz, DMSO-d₆) δ 7.76 (s, 1H), 7.75 (s, 1H),4.76 (s, 2H), 3.93 (s, 3H).

Synthesis of 5-bromo-7-fluoro-2-(4-methoxybenzyl) isoindolin-1-one (3)

A solution of methyl 4-bromo-2-(bromomethyl)-6-fluorobenzoate (2, 2.2 g,6.79 mmol), 4-methoxybenzylamine (1.87 g, 13.58 mmol) and triethylamine(2.06 g, 20.37 mmol) in N,N-dimethylformamide (20 ml) was allowed tostir at room temperature for 48 h. Progress of the reaction wasmonitored by TLC. After completion, the reaction mass was quenched withwater and the aqueous solution was extracted twice with ethyl acetate.The ethyl acetate layer was washed with cold water then cold brine. Theethyl acetate layer was separated, dried over sodium sulphate andconcentrated under reduced pressure. The residue was purified via columnchromatography using 26% ethyl acetate in hexane to get5-bromo-7-fluoro-2-(4-methoxybenzyl) isoindolin-1-one (3) as a yellowsolid. Yield: 1.7 g, 72%; MS (ESI) m/z 350.03[M+1]⁺; ¹H NMR (400 MHz,DMSO-d₆) δ 7.65 (s, 1H), 7.62 (s, 1H), 7.22 (d, J=8.4 Hz, 2H), 6.92 (d,J=8.4 Hz, 2H), 4.60 (s, 2H), 4.33 (s, 2H), 3.72 (s, 3H).

Synthesis of 7-fluoro-2-(4-methoxybenzyl)-5-(7H-pyrrolo [2, 3-d]pyrimidin-7-yl) isoindolin-1-one (5)

The synthesis of intermediate 5 was carried out according to the generalprotocol described above in Procedure C. Off-white solid; Yield: 0.28 g,51%; MS (ESI) m/z 389.19[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.16 (s,1H), 8.94 (s, 1H), 8.16 (d, J=4.0 Hz, 1H), 8.08 (s, 1H), 8.03 (d, J=10.8Hz, 1H), 7.25 (d, J=8.0 Hz, 2H), 6.95 (t. J=4.0 Hz, 2H), 6.92 (s, 1H),4.65 (s, 2H), 4.47 (s, 2H), 3.74 (s, 3H).

Synthesis of 7-fluoro-5-(7H-pyrrolo [2, 3-d] pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 23)

7-fluoro-2-(4-methoxybenzyl)-5-(7H-pyrrolo [2, 3-d] pyrimidin-7-yl)isoindolin-1-one (5, 0.27 g, 0.69 mmol) was dissolved in a mixture oftrifluoroacetic acid (5 ml), triflic acid (5 ml) and dichloromethane (5ml). The reaction was stirred at 60° C. for 3 h. Progress of thereaction was monitored by TLC. After completion, the reaction mass wasquenched with water and washed with ethyl acetate. The resulting aqueouslayer was basified with saturated aqueous sodium bicarbonate andextracted with ethyl acetate twice. The organic layer was again washedwith brine, separated, dried over sodium sulphate and concentrated underreduced pressure. The residue was purified via column chromatographyusing 2% methanol in dichloromethane to get 7-fluoro-5-(7H-pyrrolo [2,3-d] pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 23) as an off-whitesolid. Yield: 0.06 g, 32%; MS (ESI) nm/z 269.05[M+1]⁺; ¹H NMR (400 MHz,DMSO-d₆) δ 9.17 (s, 1H), 8.95 (s, 1H), 8.69 (s, 1H), 8.21 (d, J=3.6 Hz,1H), 8.14 (s, 1H), 7.99 (d, J=11.2 Hz, 1H), 6.96 (d, J=3.6 Hz, 1H), 4.50(s, 2H).

Example 24 Synthesis of 5-(5-(2-chlorophenyl)-7H-pyrrolo [2, 3-d]pyrimidin-7-yl) isoindolin-1-one (Cpd. No. 24)

Synthesis of 5-(5-(2-chlorophenyl)-7H-pyrrolo [2, 3-d] pyrimidin-7-yl)isoindolin-1-one (Cpd. No. 24)

The synthesis of compound 24 was carried out according to the generalprotocol described above in Procedure D. White solid; Yield: 0.04 g,18%; MS (ESI) m/z 361.011[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.10 (s,1H), 8.99 (s, 1H), 8.71 (s, 1H), 8.39 (s, 1H), 8.24 (s, 1H), 8.11 (d,J=6.8 Hz, 1H), 7.88 (d, J=8.0 Hz, 1H), 7.75 (d, J=8.0 Hz, 1H), 7.68 (d,J=8.0 Hz, 1H), 7.49 (m, 2H), 4.51 (s, 2H).

Example 25 Synthesis of5-(5-(thiazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(Cpd. No. 25)

Synthesis of2-(4-methoxybenzyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(3)

The synthesis of intermediate 3 was carried out according to the generalprotocol described above in Procedure C. Yellow solid. Yield: 3.0 g,54%. MS (ESI) m/z 371.2[M+1]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 9.15 (s, 1H),8.90 (s, 1H), 8.12-8.10 (m, 2H), 8.07 (d, J=8.4 Hz, 1H), 7.90 (d, J=8.0Hz, 1H), 7.24 (d, J=8.4 Hz, 2H), 6.92 (m, 3H), 4.69 (s, 2H), 4.44 (s,2H), 3.73 (s, 3H).

Synthesis of5-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-(4-methoxybenzyl)isoindolin-1-one(4)

Small portions of N-bromosuccinimide (1.15 g, 6.47 mmol) were added to astirring solution of2-(4-methoxybenzyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(3, 2 g, 5.39 mmol) in N,N-dimethylformamide (30 mL), at roomtemperature. The reaction mass was stirred at room temperature for 1 h.After completion, the reaction mixture was diluted with water (50 mL)and extracted with ethyl acetate (2×50 mL). The organic layers werecombined, dried using magnesium sulfate, filtered and concentrated todryness under vacuum. The crude was then purified by flash columnchromatography using 2.5% methanol in dichloromethane as the eluant. Thedesired fractions were concentrated to dryness under vacuum to afford5-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-(4-methoxybenzyl)isoindolin-1-one(4) as a brown solid. Yield: 1.2 g, 49%. MS (ESI) m/z 451.19[M+1]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 9.07 (s, 1H), 8.98 (s, 1H), 8.41 (s, 1H), 8.09(s, 1H), 8.03 (d, J=7.2 Hz, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.23 (d, J=8.4Hz, 2H), 6.92 (d, J=8.4 Hz, 2H), 4.69 (s, 2H), 4.43 (s, 2H), 3.73 (s,3H).

Synthesis of2-(4-methoxybenzyl)-5-(5-(thiazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(6)

The synthesis of intermediate 6 was carried out according to the generalprotocol described above in Procedure D. Yellow solid; Yield: 0.35 g,50%. MS (ESI) m/z 354.22[M+1]⁺.

Synthesis of5-(5-(thiazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(Cpd. No. 25)

A solution containing2-(4-methoxybenzyl)-5-(5-(thiazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one(0.35 g, 0.77 mmol) in trifluoroacetic acid (5 mL), triflic acid (5 mL)and dichloromethane (5 mL) was heated at 60° C. for 16 h. Aftercompletion, the reaction mixture was concentrated to dryness, quenchedwith an aqueous solution of sodium bicarbonate until the pH is 8.0 andextracted with 10% methanol in dichloromethane (2×50 mL). The organiclayers were combined, dried using magnesium sulfate and concentrated todryness under vacuum. The crude was then purified by prep HPLC and thedesired fractions were concentrated to dryness under vacuum to afford5-(5-(thiazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)isoindolin-1-one asa yellow solid. Yield: 0.020 g, 8%. MS (ESI) m/z 334.05[M+1]⁺; ¹H NMR(400 MHz, DMSO-d₆) δ 9.69 (s, 1H), 9.31 (s, 1H), 8.99 (s, 1H), 8.69 (s,2H), 8.25 (m, 2H), 8.11 (d, J=8.4 Hz, 1H), 7.88 (d, J=8.0 Hz, 1H), 4.50(s, 2H).

Example 26 Synthesis of2-(7-(1-oxoisoindolin-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)benzonitrile(Cpd. No. 26)

Synthesis of 2-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)benzonitrile (3)

The synthesis of intermediate 3 was carried out according to the generalprotocol described above in Procedure D. Yellow solid. Yield: 0.75 g,crude. MS (ESI) m/z: 221 [M+1]⁺. LCMS: 44%

Synthesis of2-[7-(1-oxoisoindolin-5-yl)pyrrolo[2,3-d]pyrimidin-5-yl]benzonitrile(Cpd. No. 26)

The synthesis of compound 26 was carried out according to the generalprotocol described above in Procedure C. White solid; Yield: 0.025 g,5%. MS (ESI) m/z 352.2[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.25 (s, 1H),9.02 (s, 1H), 8.73 (s, 1H), 8.57 (s, 1H), 8.27 (s, 1H), 8.07-8.04 (m,2H), 7.96-7.85 (m, 3H), 7.65-7.61 (m, 1H), 4.52 (s, 2H).

Example 27 Synthesis of4′-chloro-6′-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclopentane-1,1′-isoindolin]-3′-one(Cpd. No. 27)

Synthesis of6′-bromo-4′-chloro-2′-(4-methoxybenzyl)spiro[cyclopentane-1,1′-isoindolin]-3′-one (3)

To a solution of5-bromo-7-chloro-2-[(4-methoxyphenyl)methyl]isoindolin-1-one (1.0.4 g,1.09 mmol) in tetrahydrofuran (25 mL) at room temperature was addedsodium hydride (131 mg, 5.45 mmol). The reaction was stirred for 30 minand then 1,4-diiodobutane (2, 1691 mg, 5.45 mmol) was added to thereaction mixture. The reaction was stirred at room temperature for anadditional 5 h. After completion, the reaction mass was quenched with acold saturated solution of ammonium chloride at 0° C. The residue wasdissolved in ethyl acetate (100 mL) and the organic layer was washedwith water (2×20 mL) then with brine (10 mL). The organics wereseparated and dried using magnesium sulfate before concentration todryness. The crude was then purified by flash column chromatographyusing 10% ethyl acetate in hexane as the eluant. The desired fractionswere concentrated to dryness under vacuum to afford5′-bromo-7′-chloro-2′-[(4-methoxyphenyl)methyl]spiro[cyclopentane-1,3′-isoindoline]-1-oneas a yellow solid. Yield: 0.21 g, 45%; MS (ESI) m/z 422.2[M+1]⁺; ¹H NMR(400 MHz. CDCl₃) δ 7.54 (s, 1H), 7.40 (s, 1H), 7.26 (d, J=8.10 Hz, 2H),6.83 (d, J=8.10 Hz, 2H), 4.64 (s, 2H), 3.95 (s, 3H), 2.17-1.72 (m, 8H).

Synthesis of4′-chloro-2′-(4-methoxybenzyl)-6′-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclopentane-1,1′-isoindolin]-3′-one(5)

The synthesis of intermediate 5 was carried out according to the generalprotocol described above in Procedure C. Brown solid: Yield: 0.065 g,29%; ¹H NMR (400 MHz. DMSO-d₆) δ 9.16 (s, 1H), 8.95 (s, 1H), 8.30-8.27(m, 2H), 8.11 (s, 1H), 7.28 (d, J=8.04 Hz, 2H), 6.95 (d, J=3.28 Hz, 1H),6.90 (d, J=8.52 Hz, 2H), 4.64 (s, 2H), 3.71 (s, 3H), 2.01-1.92 (m, 8H).

Synthesis of4′-chloro-6′-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclopentane-1,1′-isoindolin]-3′-one(Cpd. No. 27)

Procedure F: A solution of7′-chloro-2′-[(4-methoxyphenyl)methyl]-5′-pyrrolo[2,3-d]pyrimidin-7-yl-spiro[cyclopentane-1,3′-isoindoline]-1′-one(5, 0.06 g, 0.13 mmol) in dichloromethane (5 mL) and trifluoroaceticacid (10 mL) was heated at 60° C. for 48 h. After completion, thereaction mixture was cooled to room temperature and concentrated. Thecrude was co-evaporated with dichloromethane and then liquid ammonia wasadded to neutralize the reaction mass. The crude was then purified byflash column chromatography using a gradient (2-10%)methanol indichloromethane. The desired column fractions were concentrated todryness under vacuum to afford4′-chloro-6′-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclopentane-1,1′-isoindolin]-3′-one(Cpd. No. 27) as a brown solid. Yield: 0.025 g, 56%; MS (ESI) m/z338.87[M+1]⁺; ¹H NMR (400 MHz. DMSO-d₆) δ 9.16 (d, J=8.0 Hz, 2H), 8.96(s, 1H), 8.25 (d, J=6.48 Hz, 2H), 8.13 (d, J=1.5 Hz, 1H), 6.96 (d, J=3.8Hz, 1H), 2.19-2.16 (m, 2H), 1.93 (m, 4H), 1.81-1.78 (m, 2H).

Example 28 Synthesis of6′-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4′-chlorospiro[cyclohexane-1,1′-isoindolin]-3′-one(Cpd. No. 28)

Synthesis of5′-bromo-7′-chloro-2′-[(4-methoxyphenyl)methyl]spiro[cyclohexane-1,3′-isoindoline]-1′-one(3)

To a solution of5-bromo-7-chloro-2-[(4-methoxyphenyl)methyl]isoindolin-1-one (1, 2.0 g,5.45 mmol) in tetrahydrofuran (25 mL) at room temperature was addedsodium hydride (654 mg, 27.27 mmol). The reaction was stirred for 30 minand then 1,5-diiodopentane (2, 8835 mg, 27.27 mmol) was added to thereaction mixture. After stirring at room temperature for 5 h, hereaction mass was quenched with a cold solution of saturated ammoniumchloride solution at 0° C. The residue was dissolved in ethyl acetate(100 mL) and the organic layer was washed with water (2×20 mL) then withbrine solution (10 mL). The organic layers were separated and driedusing magnesium sulfate, filtered and concentrated. The crude was thenpurified by flash column chromatography eluting with 10% ethyl acetatein hexane. The desired fractions were concentrated to dryness undervacuum to afford 5′-bromo-7′-chloro-2′-[(4-methoxyphenyl)methyl]spiro[cyclohexane-1,3′-isoindoline]-1′-one (3) as a yellow solid.Yield: 1.4 g, 60%. MS (ESI) m/z 436.44[M+1]⁺; ¹H NMR (400 MHz. DMSO-d₆)δ 8.06 (s, 1H), 7.84 (s, 1H), 7.23 (d, J=8.10 Hz, 2H), 6.87 (d, J=8.10Hz, 2H), 4.64 (s, 2H), 3.72 (s, 3H), 1.98-1.90 (m, 3H), 1.78-1.71 (m,5H), 1.49-1.40 (m, 2H).

Synthesis of6′-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4′-chloro-2′-(4-methoxybenzyl)spiro[cyclohexane-1,1′-isoindolin]-3′-one(5)

The synthesis of intermediate 5 was carried out according to the generalprotocol described above in Procedure C, brown coloured solid. Yield:0.08 g, 29%. MS (ESI) m/z 488.59 [M+1⁺]; LCMS: 89%

Synthesis of6′-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4′-chlorospiro[cyclohexane-1,1′-isoindolin]-3′-one(Cpd. No. 28)

The synthesis of compound 28 was carried out according to the generalprotocol described above in Procedure F. Brown solid; Yield: 0.025 g,55%; MS (ESI) m/z 368.33[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.32 (s,1H), 8.25 (s, 1H), 8.20 (s, 1H), 8.08 (s, 1H) 7.71 (d, J=3.72 Hz, 1H),7.24 (s, 2H), 6.84 (d, J=3.72 Hz, 1H), 2.01 (s, 2H), 1.70 (m, 5H),1.43-1.40 (m, 3H).

Example 29 Synthesis of4′-chloro-6′-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclohexane-1,1′-isoindolin]-3′-one(Cpd. No. 29)

Synthesis of4′-chloro-2′-(4-methoxybenzyl)-6′-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclohexane-1,1′-isoindolin]-3′-one(3)

The synthesis of intermediate 3 was carried out according to the generalprotocol described above in Procedure C. Brown solid; Yield: 0.18 g,64%; MS (ESI) m/z 473.4[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.16 (s, 1H),8.93 (s, 1H), 8.27 (s, 1H), 8.16 (s, 1H), 7.34-7.26 (m, 2H), 6.94 (s,1H), 6.88-6.86 (m, 2H), 6.57 (bs, 1H), 4.69 (s, 2H), 3.71 (s, 3H),1.98-1.93 (m, 4H), 1.82-1.75 (m, 3H), 1.41-1.39 (s, 3H).

Synthesis of4′-chloro-6′-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[cyclohexane-1,1′-isoindolin]-3′-one(Cpd. No. 29)

The synthesis of compound 29 was carried out according to the generalprotocol described above in Procedure F. Brown solid; Yield: 0.050 g,45%; MS (ESI) n/z 352.87[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.39 (s,1H), 9.16 (s, 1H), 8.96 (s, 1H), 8.25-8.23 (m, 2H), 8.16 (d, J=1.6 Hz,1H), 6.96 (d, J=3.72 Hz, 1H), 2.05-1.97 (m, 2H), 1.70 (m, 5H), 1.45-1.43(m, 3H).

Example 30 Synthesis of6′-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4′-chlorospiro[cyclopentane-1,1′-isoindolin]-3′-one(Cpd. No. 30)

Synthesis of6′-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4′-chloro-2′-(4-methoxybenzyl)spiro[cyclopentane-1,1′-isoindolin]-3′-one(3)

The synthesis of intermediate 3 was carried out according to the generalprotocol described above in Procedure C. Brown solid; Yield: 0.20 g,crude; MS (ESI) m/z 474.35[M+1]⁺.

Synthesis of6′-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4′-chlorospiro[cyclopentane-1,1′-isoindolin]-3′-one(Cpd. No. 30)

The synthesis of compound 30 was carried out according to the generalprotocol described above in Procedure F. Brown solid; Yield: 0.020 g,55%; MS (ESI) m/z 358.28[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.08 (s,1H), 8.23 (s, 1H), 8.19 (s, 1H), 8.05 (s, 1H) 7.79 (d, J=3.52 Hz, 1H),7.24 (s, 2H), 6.84 (d, J=3.6 Hz, 1H), 2.17-1.77 (m, 8H).

Example 31 Synthesis of 6-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1′,4-dimethylspiro[isoindoline-1,4′-piperidin]-3-one(Cpd. No. 31)

Synthesis of 6-(4-amino-7H-pyrrolo [2, 3-d]pyrimidin-7-yl)-2-(4-methoxybenzyl)-1,4-dimethylspiro[isoindoline-1,4′-piperidin]-3-one(3)

The synthesis of intermediate 3 was carried out according to the generalprotocol described above in Procedure C. Light brown solid; Yield: 0.25g, crude; MS (ESI) m/z 483[M+1]⁺.

Synthesis of 6-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1,4′-dimethylspiro[isoindoline-1,4′-piperidin]-3-one(Cpd. No. 31)

The synthesis of compound 31 was carried out according to the generalprotocol described above in Procedure F. Off white solid; Yield: 30 mg,16%; MS (ESI) m/z 363.19[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.14 (s,1H), 8.16 (s, 1H), 7.93 (s, 1H), 7.78 (s, 1H), 7.69 (d, J=3.7 Hz, 1H),7.19 (s, 2H), 6.80 (d, J=3.7 Hz, 1H), 2.75-2.85 (m, 2H), 2.65 (s, 3H),2.40-2.17 (m, 7H), 1.41 (m, 2H).

Example 32 Synthesis of2-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one(Cpd. No. 32)

Synthesis of 6-(4-methoxybenzyl)-2-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one (3)

The synthesis of intermediate 3 was carried out according to the generalprotocol described above in Procedure C. Off white solid; Yield: 0.060g, 16%; MS (ESI) m/z 372.11[M+1]⁺.

Synthesis of 2-(7H-pyrrolo[2, 3-d]pyrimidin-7-yl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-5-one (Cpd. No. 32)

The synthesis of compound 32 was carried out according to the generalprotocol described above in Procedure F. Off white solid; Yield: 0.040g, 55%; MS (ESI) m/z 252.07[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆ with TFA-d)δ 9.54 (s, 1H), 9.36 (s, 1H), 8.74-8.72 (m, 2H), 8.34 (d, J=8.4 Hz, 1H),7.22 (d, J=4.0 Hz, 1H), 4.45 (s, 2H).

Example 33 Synthesis of6-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1′-(2,2-difluoroethyl)-4-methylspiro[isoindoline-1,4′-piperidin]-3-one(Cpd. No. 33)

Synthesis of 6-(4-amino-7H-pyrrolo [2,3-d]pyrimidin-7-yl)-1′-(2,2-difluoroethyl)-2-(4-methoxybenzyl)-4-methylspiro[isoindoline-1,4′-piperidin]-3-one(3)

The synthesis of intermediate 3 was carried out according to the generalprotocol described above in Procedure C. Brown solid; Yield: 0.35 g,crude; MS (ESI) m/z 532.24[M+1]⁺.

Synthesis of6-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1′-(2,2-difluoroethyl)-4-methylspiro[isoindoline-1,4′-piperidin]-3-one(Cpd. No. 33)

The synthesis of compound 33 was carried out according to the generalprotocol described above in Procedure F. White solid; Yield: 7 mg, 3%,MS (ESI) m/z 412.18[M+1]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 9.162 (s, 1H),8.17 (s, 1H), 7.95 (s, 1H), 7.83 (s, 1H), 7.70 (d, J=3.72 Hz, 1H), 7.16(s, 2H), 6.80 (d, J=3.64 Hz, 1H), 6.31-6.03 (tt, J=55.8, 3.46 Hz, 1H)2.92-2.87 (m, 2H), 2.86-2.77 (m, 2H), 2.71-2.63 (m, 2H), 2.66 (s, 3H),2.23-2.18 (m, 2H), 1.40-1.33 (m, 2H).

Example 34 Synthesis of2′-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4′-methylspiro[cyclohexane-1,7′-pyrrolo[3,4-b]pyridin]-5′(6′H)-one(Cpd. No. 34)

Procedure G: To a solution of 7H-pyrrolo[2,3-d]pyrimidin-4-amine (1,0.32 g, 2.39 mmol) and2′-chloro-4′-methylspiro[cyclohexane-1,7′-pyrrolo[3,4-b]pyridin]-5′(6′H)-one(2, 0.6 g, 2.39 mmol) in 1,4-dioxane (15 mL) was added cesium carbonate(2.33 g, 7.17 mmol). The reaction mixture was purged with argon for 5min, and then XanthPhos (69 mg, 0.11 mmol). XPhos (57 mg, 0.11 mmol),tris(dibenzylidcneacetone)dipalladium(0) (109 mg, 0.11 mmol) andpalladium acetate (27 mg, 0.11 mmol) were added and the reaction mixturepurged for an additional 5 min. The purged reaction mixture was stirredat 100° C. for 4 h. After TLC showed completion, the reaction mixturewas filtered through a bed of celite and the resulting filtrate wasconcentrated. The crude product was purified by preparative HPLC. Thedesired fractions were concentrated to dryness under vacuum to afford2′-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4′-methylspiro[cyclohexane-1,7′-pyrrolo[3,4-b]pyridin]-5′(6′H)-oneas a yellow solid. Yield: 0.095 g, 11%; MS (ESI) nm/z 348.4[M+1]⁺; ¹HNMR: (400 MHz, DMSO-d₆) δ 12.36-12.28 (bs, 1H), 11.04-10.90 (bs, 1H),9.12 (s, 1H), 8.64 (s, 1H), 7.10 (bs, 1H), 7.46 (s, 1H), 7.14 (s, 1H),2.62 (s, 3H), 2.11-2.06 (m, 2H), 1.72-1.68 (m, 5H), 1.41-1.38 (m, 3H).

Example 35 Synthesis of7-(3′,4′-dimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(Cpd. No. 35)

Synthesis of6′-methoxy-4′-methyl-3′-methylene-2′,3′-dihydrospiro[cyclohexane-1,1′-indene](2)

To a suspension of potassium tert-butoxide (1.75 g, 15.60 mmol) intetrahydrofuran (20 mL) is added methyltriphenylphosphonium bromide(5.46 g, 15.28 mmol). The reaction is stirred at room temperature for 1h and then cooled to 0° C. A solution of6′-methoxy-4′-methylspiro[cyclohexane-1,1′-inden]-3′(2′H)-one (1, 3.15g, 12.89 mmol) in tetrahydrofuran (10 mL) is added. The mixture isstirred at room temperature for 16 h, poured into water and extractedwith ethyl acetate. The organic phase is dried over magnesium sulfate,filtered and concentrated. Purification via column chromatographyaffords6′-methoxy-4′-methyl-3′-methylene-2′,3′-dihydrospiro[cyclohexane-1,1′-indene](2).

Synthesis of 6′-methoxy-3,4′-dimethyl-2,3′-dihydrospiro[cyclohexane-1′-indene] (3)

To a solution of6′-methoxy-4′-methyl-3′-methylene-2′,3′-dihydrospiro[cyclohexane-1,1′-indene](2, 1.00 g, 4.13 mmol) in ethanol (20 mL) is added 10% palladium oncarbon (100 mg). The reaction is purged with hydrogen and stirred atroom temperature overnight. The mixture is filtered through a pad ofcelite, concentrated and purified via column chromatography to afford6′-methoxy-3′,4′-dimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-indene](3).

Synthesis of3,4′-dimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-ol (4)

To a solution of6′-methoxy-3′,4′-dimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-indene](3,1.00 g, 4.09 mmol) in dichloromethane (20 mL) at −78° C. is added slowlyboron tribromide (0.79 mL, 8.18 mmol). The reaction is allowed to stirat room temperature for 16 h. After completion, the reaction mixture isquenched with saturated aqueous sodium bicarbonate solution to adjust topH 8. The mixture is extracted with dichloromethane (2×30 mL). Thecombined organics is dried over sodium sulfate, filtered andconcentrated. The crude is then purified via column chromatography toafford 3′,4′-dimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-ol(4).

Synthesis of3,4′-dimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-yltrifluoromethanesulfonate (5)

To a solution of3′,4′-dimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-ol (4, 1.00g, 4.34 mmol) in dichloromethane (15 mL) at −30° C.,diisopropylethylamine (1.28 mL, 7.38 mmol) is added followed by the slowaddition of triflic anhydride (0.80 mL, 4.77 mmol). The reaction isallowed to stir at room temperature for 1 h. After completion, thereaction mixture is basified by saturated aqueous sodium bicarbonatesolution to pH 8. The mixture is extracted with dichloromethane (2×10mL). The combined organics is dried over sodium sulfate, filtered andconcentrated to dryness under vacuum. The crude is then purified viacolumn chromatography to afford3′,4′-dimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-yltrifluoromethanesulfonate (5).

Synthesis of7-(3′,4′-dimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(Cpd. No. 35)

The synthesis of compound 35 is carried out as described above using thegeneral protocol of Procedure G.

Example 36 Synthesis of7-(3′,3′,4′-trimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(Cpd. No. 36)

Synthesis of methyl 2-(4-methoxy-2-methylphenyl)-2-methylpropanoate (2)

To a solution of methyl 2-(4-methoxy-2-methylphenyl)acetate (1, 1.00 g,5.15 mmol) in tetrahydrofuran (20 mL) at 0° C., sodium hydride (0.31 g,12.88 mmol) is added portion wise and the reaction mixture is allowed tostir at room temperature for 30 min. Iodomethane (0.96 mL, 15.45 mmol)is added and the reaction mixture is allowed to stir at 70° C. for 16 h.The reaction mixture is quenched with water and is extracted in ethylacetate. The organic layer is separated, dried over sodium sulphate,filtered and concentrated under reduced pressure. The residue ispurified by silica gel column chromatography to afford methyl2-(4-methoxy-2-methylphenyl)-2-methylpropanoate (2).

Synthesis of 2-(4-methoxy-2-methylphenyl)-2-methylpropanoic acid (3)

To a solution of methyl 2-(4-methoxy-2-methylphenyl)-2-methylpropanoate(2, 1.00 g, 4.50 mmol) in tetrahydrofuran (10 mL) and ethanol (10 mL) isadded 1 M lithium hydroxide aqueous solution (10 mL). The reaction isstirred at room temperature overnight. The mixture is diluted with waterand extracted with ethyl acetate. The organic layer is washed withbrine, dried over sodium sulfate, filtered and concentrated. The crudeobtained is further purified via column chromatography to obtain2-(4-methoxy-2-methylphenyl)-2-methylpropanoic acid (3).

Synthesis of 1-diazo-3-(4-methoxy-2-methylphenyl)-3-mnethylbutan-2-one(4)

To a solution of 2-(4-methoxy-2-methylphenyl)-2-methylpropanoic acid (3,1.00 g, 4.80 mmol) in dichloromethane (10 mL) at 0° C. is added oxalylchloride (1 M in dichloromethane, 5.28 mL, 5.28 mmol) followed by twodrops of N,N-dimethylformamide. The reaction is stirred at roomtemperature for 1 h. The mixture is concentrated and dried under vacuum.The residue is dissolved in dichloromethane (10 mL). To this solution at0° C. is purged with diazomethane. The reaction is fitted with a calciumchloride drying tube and allowed to stand at room temperature for 16 h.The mixture is purged with nitrogen and concentrated. The residue ispurified via column chromatography to afford1-diazo-3-(4-methoxy-2-methylphenyl)-3-methylbutan-2-one (4).

Synthesis of 5-methoxy-1, 1,7-trimethyl-1,3-dihydro-2H-inden-2-one (5)

To a solution of1-diazo-3-(4-methoxy-2-methylphenyl)-3-methylbutan-2-one (4, 1.00 g,4.30 mmol) in dichloromethane (10 mL) is added rhodium (II) acetatedimer dihydrate (105 mg, 0.22 mmol). The reaction is stirred at roomtemperature overnight. The mixture is filtered through a pad of celite,concentrated and purified via column chromatography to afford5-methoxy-1,1,7-trimethyl-1,3-dihydro-2H-inden-2-one (5).

Synthesis of6′-methoxy-3,3,4′-trimethylspiro[cyclohexane-1,1′-inden]-2′(3′H)-one (7)

To a solution of 5-methoxy-1,1,7-trimethyl-1,3-dihydro-2H-inden-2-one(5, 1.00 g, 4.90 mmol) in tetrahydrofuran (20 mL) at 0° C., sodiumhydride (0.29 g, 12.25 mmol) is added portion wise and the reactionmixture is allowed to stir at room temperature for 30 min.1,5-Dibromopentane (6, 1.13 g, 4.9 mmol) is added and the reactionmixture is allowed to stir at 70° C. for 16 h. The reaction mixture isquenched with water and is extracted in ethyl acetate. The organic layeris separated, dried over sodium sulphate, filtered and concentratedunder reduced pressure. The residue is purified by silica gel columnchromatography to afford6′-methoxy-3′,3′,4′-trimethylspiro[cyclohexane-1,1′-inden]-2′(3′H)-one(7).

Synthesis of6′-methoxy-3′,3,4′-trimethyl-2,3′-dihydrospiro[cyclohexane-1,1′-indene](8)

To a solution of6′-methoxy-3′,3′,4′-trimethylspiro[cyclohexane-1,1′-inden]-2′(3′H)-one(7, 1.00 g, 3.67 mmol) in ethylene glycol (40 mL) is added hydrazinehydrate solution (78-82%, 0.25 g, 4.04 mmol) followed by potassiumhydroxide (0.62 g, 11.01 mmol). The reaction is fitted with a Dean-Starktrap and stirred at 120° C. for 3 h to distill off water and excesshydrazine. The reaction is then stirred at reflux overnight. The mixtureis cooled to room temperature, diluted with water and extracted withethyl acetate. The combined organics is dried over magnesium sulfate,filtered and concentrated. The crude is purified via columnchromatography to afford6′-methoxy-3′,3′,4′-trimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-indene](8).

Synthesis of3′,3′,4′-trimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-ol (9)

To a solution of6′-methoxy-3′,3′,4′-trimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-indene](8,1.00 g, 3.87 mmol) in dichloromethane (20 mL) at −78° C. is added slowlyboron tribromide (0.74 mL, 7.74 mmol). The reaction is allowed to stirat room temperature for 16 h. After completion, the reaction mixture isquenched with saturated aqueous sodium bicarbonate solution to adjust topH 8. The mixture is extracted with dichloromethane (2×30 mL). Thecombined organics is dried over sodium sulfate, filtered andconcentrated. The crude is then purified via column chromatography toafford3′,3′,4′-trimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-ol (9).

Synthesis of 3,3′,4′-trimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-yl trifluoromethanesulfonate(10)

To a solution of3′,3′,4′-trimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-ol (9,1.00 g, 4.09 mmol) in dichloromethane (15 mL) at −30° C.diisopropylethylamine (1.21 mL, 6.95 mmol) is added followed by the slowaddition of triflic anhydride (0.76 mL, 4.50 mmol). The reaction isallowed to stir at room temperature for 1 h. After completion, thereaction mixture is basified by saturated aqueous sodium bicarbonatesolution to pH 8. The mixture is extracted with dichloromethane (2×20mL). The combined organics is dried over sodium sulfate, filtered andconcentrated to dryness under vacuum. The crude is then purified viacolumn chromatography to afford3′,3′,4′-trimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-yltrifluoromethanesulfonate (10).

Synthesis of7-(3′,3′,4′-trimethyl-2′,3′-dihydrospiro[cyclohexane-1,1′-inden]-6′-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(Cpd. No. 36)

The synthesis of compound 36 is carried out as described above using thegeneral protocol of Procedure G.

Example 37 Synthesis of7-(3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-6′-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(Cpd. No. 37)

Synthesis of 6′-methoxy-4′-methylspiro[cyclohexane-1,1′-isoindoline] (2)

To a solution of6′-methoxy-4′-methylspiro[cyclohexane-1,1′-isoindolin]-3′-one (1, 1.00g, 4.08 mmol) in tetrahydrofuran (20 mL) is added dropwise boranedimethyl sulfide complex (12.24 mL, 24.48 mmol, 2 M in tetrahydrofuran).The reaction is stirred at 65° C. for 7 h, then stirred at roomtemperature overnight, 0.5 M hydrochloric acid (8 mL) is added dropwiseand the mixture is refluxed for 2 h. The mixture is cooled to roomtemperature, basified with 1 M aqueous sodium hydroxide solution to pH=8and extracted with ethyl acetate. The combined organics is dried overmagnesium sulfate, filtered and concentrated. The crude is purified viacolumn chromatography to afford methyl6′-methoxy-4′-methylspiro[cyclohexane-1,1′-isoindoline] (2).

Synthesis ofN-tert-butyl-1-(6′-methoxy-4′-methylspiro[cyclohexane-1,1′-isoindolin]-2′-yl)methanimine(4)

To a solution of methyl6′-methoxy-4′-methylspiro[cyclohexane-1,1′-isoindoline] (2, 1.00 g, 4.32mmol) in toluene (20 mL) is added ammonium sulfate (1.14 g, 8.64 mmol)followed by N′-tert-butyl-N,N-dimethylformimidamide (3, 0.83 g, 6.48mmol). The reaction is refluxed overnight. The mixture is cooled to roomtemperature, filtered and concentrated. The crude is purified via columnchromatography to affordN-tert-butyl-1-(6′-methoxy-4′-methylspiro[cyclohexane-1,1′-isoindolin]-2′-yl)methanimine(4).

Synthesis ofN-tert-butyl-1-(6′-methoxy-3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-2′-yl)methaninine(5)

To a solution ofN-tert-butyl-1-(6′-methoxy-4′-methylspiro[cyclohexane-1,1′-isoindolin]-2′-yl)methanimine(4, 1.00 g, 3.18 mmol) in tetrahydrofuran (20 mL) at −78° C. is addedn-butyl lithium (1.6 M in hexanes, 2.19 mL, 3.50 mmol) dropwise and thereaction is stirred for 30 min. Iodomethane (0.30 mL, 4.77 mmol) isadded and the reaction is warmed to room temperature and stirred for 1h. The reaction mixture is quenched with water and extracted with ethylacetate. The combined organics is dried over magnesium sulphate,filtered and concentrated. The residue is purified via columnchromatography toN-tert-butyl-1-(6′-methoxy-3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-2′-yl)methanimine(5).

Synthesis of 3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-6′-ol (6)

To a solution ofN-tert-butyl-1-(6′-methoxy-3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-2′-yl)methanimine(5, 1.00 g, 3.04 mmol) in dichloromethane (20 mL) at −78° C. is addedslowly boron tribromide (0.59 mL, 6.08 mmol). The reaction is stirred atroom temperature for 16 h. After completion, the reaction mixture isquenched with saturated aqueous sodium bicarbonate solution to adjust topH 8. The mixture is extracted with dichloromethane (2×30 mL). Thecombined organics is dried over sodium sulfate, filtered andconcentrated. The crude is then purified via column chromatography toafford 3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-6′-ol (6).

Synthesis of tert-butyl6′-hydroxy-3′,4′-dimethylspiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(7)

To a solution of 3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-6′-ol(6, 1.00 g, 4.32 mmol) and di-tert-butyl dicarbonate (1.19 mL, 5.18mmol) in tetrahydrofuran (20 mL) is added a solution of potassiumcarbonate (1.49 g, 10.80 mmol) in water (20 mL). The reaction is stirredat room temperature overnight. The mixture is diluted with brine andextracted with ethyl acetate. The combined organics is dried overmagnesium sulfate, filtered and concentrated. The crude is then purifiedvia column chromatography to afford tert-butyl6′-hydroxy-3′,4′-dimethylspiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(7).

Synthesis of tert-butyl3′,4′-dimethyl-6′-(((trifluoromethyl)sulfonyl)oxy)spiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(8)

To a solution of tert-butyl6′-hydroxy-3′,4′-dimethylspiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(7, 1.00 g, 3.02 mmol) in dichloromethane (15 mL) at −30° C.,diisopropylethylamine (0.89 mL, 5.13 mmol) is added followed by the slowaddition of triflic anhydride (0.56 mL, 3.32 mmol). The reaction isallowed to stir at room temperature for 1 h. After completion, thereaction mixture is basified by saturated aqueous sodium bicarbonatesolution to pH 8. The mixture is extracted with dichloromethane (2×20mL). The combined organics is dried over sodium sulfate, filtered andconcentrated to dryness under vacuum. The crude is then purified viacolumn chromatography to afford tert-butyl3′,4′-dimethyl-6′-(((trifluoromethyl)sulfonyl)oxy)spiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(8).

Synthesis of tert-butyl6′-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3′,4′-dimethylspiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(10)

The synthesis of intermediate 10 is carried out as described above usingthe general protocol of Procedure G.

Synthesis of7-(3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-6-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(Cpd. No. 37)

The synthesis of intermediate 37 is carried out as described above usingthe general protocol of Procedure C.

Example 38 Synthesis of7-(3′,3′,4′-trimethylspiro[cyclohexane-1,1′-isoindolin]-6′-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(Cpd. No. 38)

Synthesis ofN-tert-butyl-1-(6′-methoxy-3′,3′,4′-trimethylspiro[cyclohexane-1,1′-isoindolin]-2′-yl)methaninine(2)

To a solution ofN-tert-butyl-1-(6′-methoxy-3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-2′-yl)methanimine(1, 1.00 g, 3.04 mmol) in tetrahydrofuran (20 mL) at −78° C. is addedn-butyl lithium (1.6 M in hexanes, 2.09 mL, 3.34 mmol) dropwise and thereaction is stirred for 30 min. Iodomethane (0.28 mL, 4.56 mmol) isadded and the reaction is warmed to room temperature and stirred for 1h. The reaction mixture is quenched with water and extracted with ethylacetate. The combined organics is dried over magnesium sulphate,filtered and concentrated. The residue is purified via columnchromatography toN-tert-butyl-1-(6′-methoxy-3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-2′-yl)methanimine(2).

Synthesis of 3′,3′,4′-trimethylspiro[cyclohexane-1,1′-isoindolin]-6′-ol(3)

To a solution ofN-tert-butyl-1-(6′-methoxy-3′,4′-dimethylspiro[cyclohexane-1,1′-isoindolin]-2′-yl)methanimine(2, 1.00 g, 2.92 mmol) in dichloromethane (20 mL) at −78° C. is addedslowly boron tribromide (0.56 mL, 5.84 mmol). The reaction is stirred atroom temperature for 16 h. After completion, the reaction mixture isquenched with saturated aqueous sodium bicarbonate solution to adjust topH 8. The mixture is extracted with dichloromethane (2×30 mL). Thecombined organics is dried over sodium sulfate, filtered andconcentrated. The crude is then purified via column chromatography toafford 3′,3′,4′-trimethylspiro[cyclohexane-1,1′-isoindolin]-6′-ol (3).

Synthesis of tert-butyl6′-hydroxy-3,3′,4′-trimethylspiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(4)

To a solution of3′,3′,4′-trimethylspiro[cyclohexane-1,1′-isoindolin]-6′-ol (3, 1.00 g,4.08 mmol) and di-tert-butyl dicarbonate (1.12 mL, 4.90 mmol) intetrahydrofuran (20 mL) is added a solution of potassium carbonate (1.41g, 10.20 mmol) in water (20 mL). The reaction is stirred at roomtemperature overnight. The mixture is diluted with brine and extractedwith ethyl acetate. The combined organics is dried over magnesiumsulfate, filtered and concentrated. The crude is then purified viacolumn chromatography to afford tert-butyl6′-hydroxy-3′,3′,4′-trimethylspiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(4).

Synthesis of tert-butyl3′,3′4′-trimethyl-6′-(((trifluoromethyl)sulfonyl)oxy)spiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(5)

To a solution of tert-butyl6′-hydroxy-3′,3′,4′-trimethylspiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(4, 1.00 g, 2.89 mmol) in dichloromethane (15 mL) at −30° C.,diisopropylethylamine (0.86 mL, 4.91 mmol) is added followed by the slowaddition of triflic anhydride (0.54 mL, 3.18 mmol). The reaction isallowed to stir at room temperature for 1 h. After completion, thereaction mixture is basified by saturated aqueous sodium bicarbonatesolution to pH 8. The mixture is extracted with dichloromethane (2×20mL). The combined organics is dried over sodium sulfate, filtered andconcentrated to dryness under vacuum. The crude is then purified viacolumn chromatography to afford tert-butyl3′,3′,4′-trimethyl-6′-(((trifluoromethyl)sulfonyl)oxy)spiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate(5).

Synthesis of tert-butyl6′-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3′,3′,4′-trimethylspiro[cyclohexane-1,1′-isoindoline]-2′-carboxylate (7)

The synthesis of intermediate 7 is carried out as described above usingthe general protocol of Procedure G.

Synthesis of 7-(3′,3′,4′-trimethylspiro[cyclohexane-)1′-isoindolin]-6′-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (Cpd. No. 38)

The synthesis of compound 38 is carried out as described above using thegeneral protocol of Procedure C.

Example 39 Synthesis of7-(4′-methyl-2′H-dispiro[cyclohexane-1,1′-indene-3′,1″-cyclopropan]-6′-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(Cpd. No. 39)

Synthesis of6′-methoxy-4′-methyl-2′H-dispiro[cyclohexane-1,1′-indene-3′,1″-cyclopropane](2)

A 1.1 M toluene solution of diethyl zinc (15.02 mL, 16.52 mmol) is addedto a reaction vessel containing dichloromethane (20 mL) and cooled to 0°C. Trifluoroacetic acid (1.26 mL, 16.52 mmol) is added to the resultingsolution and the reaction is stirred at 0° C. for 15 min. To the cooledsolution is added diiodomethane (1.33 mL, 16.52 mmol), and the reactionis stirred for an additional 15 min at 0° C. Then, a solution of6′-methoxy-4′-methyl-3′-methylene-2′,3′-dihydrospiro[cyclohexane-1,1′-indene](1, 1.00 g, 4.13 mmol) in dichloromethane (10 mL) is added. The reactionis maintained 0° C. for another 15 min, then allowed to gradually warmto room temperature. Upon completion, the reaction is quenched with asaturated aqueous ammonium chloride solution (40 mL) and diluted withdichloromethane (40 mL). The combined organics is washed with brine (40mL), dried over magnesium sulfate, filtered and concentrated. The crudeis purified via flash chromatography to afford6′-methoxy-4′-methyl-2′H-dispiro[cyclohexane-1,1′-indene-3′,1″-cyclopropane](2).

Synthesis of4′-methyl-2′H-dispiro[cyclohexane-1,1′-indene-3′,1″-cyclopropan]-6′-ol(3)

To a solution of6′-methoxy-4′-methyl-2′H-dispiro[cyclohexane-1,1′-indene-3′,1″-cyclopropane](2, 0.75 g, 2.92 mmol) in dichloromethane (20 mL) at −78° C. is addedslowly boron tribromide (0.56 mL, 5.84 mmol). The reaction is stirred atroom temperature for 16 h. After completion, the reaction mixture isquenched with saturated aqueous sodium bicarbonate solution to adjust topH 8. The mixture is extracted with dichloromethane (2×30 mL). Thecombined organics is dried over sodium sulfate, filtered andconcentrated. The crude is then purified via column chromatography toafford4′-methyl-2′H-dispiro[cyclohexane-1,1′-indene-3′,1″-cyclopropan]-6′-ol(3).

Synthesis of4′-methyl-2′H-dispiro[cyclohexane-1,1′-indene-3′,1″-cyclopropan]-6′-yltrifluoromethanesulfonate (4)

To a solution of4′-methyl-2′H-dispiro[cyclohexane-1,1′-indene-3′,1″-cyclopropan]-6′-ol(3, 0.70 g, 2.89 mmol) in dichloromethane (15 mL) at −30° C.,diisopropylethylamine (0.86 mL, 4.91 mmol) is added followed by the slowaddition of triflic anhydride (0.54 mL, 3.18 mmol). The reaction isallowed to stir at room temperature for 1 h. After completion, thereaction mixture is basified by saturated aqueous sodium bicarbonatesolution to pH 8. The mixture is extracted with dichloromethane (2×20mL). The combined organics is dried over sodium sulfate, filtered andconcentrated to dryness under vacuum. The crude is then purified viacolumn chromatography to afford4′-methyl-2′H-dispiro[cyclohexane-1,1′-indene-3′,1″-cyclopropan]-6′-yltrifluoromethanesulfonate (4).

Synthesis of7-(4′-methyl-2′H-dispiro[cyclohexane-1,1′-indene-3′,1″-cyclopropan]-6′-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(Cpd. No. 39)

The synthesis of compound 39 is carried out as described above using thegeneral protocol of Procedure G.

Example 40: MNK Biochemical Enzymatic Assay

Compounds are screened for MNK inhibition using the ADP-Glo kinase assaykit (Promega, catalogue No. V9101). All kinase reactions are performedin Reaction Buffer E (15 mM HEPES pH7.4, 20 mM NaCl, 1 mM EGTA, 10 mMMgCl₂, 0.1 mg/ml BGG, and 0.02% Tween-20). Final MNK 1 reactionscontained 10 nM recombinant MNK1 (Life Technologies, PR9138A), 100 μMMNK substrate peptide Ac-TATKSGSTTKNR-NH2 (American Peptide Company),300 μM ATP, and varying concentrations of the inhibitory compound ofinterest. Final MNK2 reactions contained 3 nM recombinant MNK2 (LifeTechnologies, PV5607), 50 μM MNK substrate peptide Ac-TATKSGSTTKNR-NH2(American Peptide Company), 10 μM ATP, and varying concentrations of theinhibitory compound of interest. Final DMSO concentration in eachreaction is 1%.

Kinase reactions are carried out in 96-well half-area white flat-bottompolystyrene plates in a final volume of 25 μl. MNK1/2 enzymes arepre-incubated with compound and peptide substrate for 5 minutes prior tothe addition of ATP. After the addition of ATP, kinase reactions areincubated at room temperature for 40 minutes. Reactions are subsequentlystopped by the addition of 25 μl of ADP-Glo Reagent and incubating foran additional 40 minutes. The final luminescent signal used for kinaseactivity readout is produced by the addition of 45 μl of KinaseDetection Reagent (ADP-Glo kit, Promega) and incubating for 40 minutes.The luminescent signal is detected using a Victor 2 multilabel counter(Perkin Elmer) and the concentration of compound necessary to achieveinhibition of enzyme activity by 50% (IC₅₀) is calculated using signalsfrom an 8-point compound dilution series.

The results of these assays are set forth in Table 1 below. To this end,IC₅₀ values of less than 0.01 μM are labeled as “+++”, from 0.01 to 0.1μM are labeled as “++”, and greater than 0.1 to 10.0 μM are labeled as“+” (NA means “not available”).

TABLE 1 MNK Biochemical Enzymatic Assay (IC₅₀) IC₅₀ Cpd. No. Mnk1 Mnk21 + + 2 NA + 3 NA + 4 NA + 5 NA + 6 NA + 7 NA + 8 ++ ++ 9 NA − 10 NA −11 NA + 12 NA + 13 NA + 14 NA + 15 NA − 16 NA + 17 +++ ++ 18 +++ +++ 19NA + 20 NA + 21 NA + 22 ++ ++ 23 NA + 24 NA + 25 NA + 26 NA + 27 NA +++28 NA +++ 29 NA ++ 30 NA +++ 31 NA NA 32 NA NA 33 NA NA 34 NA NA

Example 41: peIF4E Signaling Cellular Assay

Phosphorylated eIF4E is assayed using the CisBio peIF4E HTRF® assay kit(CisBio, catalogue No. 64EF4PEG). Cells are plated in 96-welltissue-culture treated plate in appropriate growth medium (90 μL).Compounds (10×) are diluted using 3-fold serial dilutions in cellculture medium and added to cells. Plates are incubated for 2 hrs at 37°C. The cell supernatant is carefully removed either by aspiratingsupernatant or by flicking the plate. Immediately 50 μL of supplementedlysis buffer (IX) is added and incubated for at least 30 minutes at roomtemperature under shaking. After homogenization by pipetting up anddown, 16 μL of cell lysate is transferred from the 96-well cell-cultureplate to a 384-well small volume white plate, 4 μL of premixed antibodysolutions (vol/vol) is prepared in the detection buffer and added. Theplate is covered with a plate sealer and incubated overnight at roomtemperature. The fluorescence emissions at two different wavelengths areread (665 nm and 620 nm) on a Wallac Victor2. Emission ratios areconverted into percent inhibitions and imported into GraphPad Prismsoftware. The concentration of compound necessary to achieve inhibitionof enzyme activity by 50% (IC₅₀) is calculated using concentrationsranging from 20 μM to 0.1 nM (12-point curve). IC₅₀ values aredetermined using a nonlinear regression model available in GraphPadPrism 5.

The results of these assays are set forth in Table 2 below. To this end,IC₅₀ values of less than 0.05 μM are labeled as “+++”, from 0.05 to 1.0μM are labeled as “++”, greater than 1.0 to 100 μM are labeled as “+”,and NA means “not available”.

TABLE 2 peIF4E Signaling Cellular Assay (IC₅₀) Cpd. No. IC₅₀ 1 + 2 NA3 + 4 NA 5 NA 6 + 7 NA 8 + 9 NA 10 NA 11 + 12 NA 13 NA 14 NA 15 NA 16 NA17 ++ 18 ++ 19 NA 20 NA 21 NA 22 + 23 NA 24 NA 25 NA 26 NA 27 ++ 28 +++29 +++ 30 +++ 31 ++ 32 ++ 33 ++ 34 +++

The various embodiments described above can be combined to providefurther embodiments. All of the U.S. patents. U.S. patent applicationpublications, U.S. patent applications, foreign patents, foreign patentapplications and non-patent publications referred to in thisspecification and/or listed in the Application Data Sheet areincorporated herein by reference, in their entirety. Aspects of theembodiments can be modified, if necessary to employ concepts of thevarious patents, applications and publications to provide yet furtherembodiments.

These and other changes can be made to the embodiments in light of theabove-detailed description. In general, in the following claims, theterms used should not be construed to limit the claims to the specificembodiments disclosed in the specification and the claims, but should beconstrued to include all possible embodiments along with the full scopeof equivalents to which such claims are entitled. Accordingly, theclaims are not limited by the disclosure.

1. A compound according to Formula (I):

a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereofwherein: A¹ is —N—; A² is —N— or —CR^(5a); A³ is —N— or —CR⁶; A⁴ is —N—or —CR^(5a); A⁵ is —NR⁷ or —CR^(7a)R^(7b); A⁶ and A⁷ are —CR^(8a) when------- represents a bond, otherwise A⁶ and A⁷ are —CR^(8a)R^(8b); W¹ isO, S, NH, NO(R⁹) or CR^(9a)R^(9b); m is 1; n is 1, 2 or 3; R¹ and R²independently are —H, —NHR¹⁰, NHR¹⁰-alkylene, (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, cycloalkyl, heterocyclyl, heteroaryl,aryl, arylalkylene, cycloalkylalkylene, heterocyclylalkylene orheteroarylalkylene; or R¹ and R² together with the carbon atom to whichthey are attached form a cycloalkyl or heterocyclyl ring; R³ and R⁴independently are —H, —OH, —CN, —SR¹⁰, S(O)₂(C₁-C₈) alkyl, —C(O)NHR¹⁰,—C(O)NR¹⁰R¹⁰, —NHR¹⁰, —NR¹⁰R¹⁰, NHR¹⁰-alkylene, NR¹⁰R¹⁰-alkylene,(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, (C₁-C₈)haloalkyl,—O(C₁-C₈)alkyl, —O(C₁-C₈)haloalkyl, —O(C₁-C₈)alkyleneNHR¹⁰,—O(C₁-C₈)alkyleneNR¹⁰R¹⁰, cycloalkyl, heterocyclyl, heteroaryl, aryl,arylalkylene, cycloalkylalkylene, heterocyclylalkylene,heteroarylalkylene, alkylaminyl, alkylcarbonylaminyl,cycloalkylcarbonylaminyl, cycloalkylaminyl, or heterocyclylaminyl;R^(5a) is —H, —OH, halogen, —CN, acetyl, —(C₁-C₈)alkyl, —S(C₁-C₈)alkyl,—(C₂-C₈)alkenyl, —(C₂-C₈)alkynyl, —O(C₁-C₈)alkyl, (C₁-C₈)haloalkyl,—NHR¹⁰, —NR¹⁰R¹⁰, NHR¹⁰-alkylene, NR¹⁰R¹⁰-alkylene, or—O(C₁-C₈)haloalkyl; R^(5b) and R⁶ is —H, —OH, —SH, —CN, —S(O)₂R¹⁰,halogen, —S(C₁-C₈)alkyl, —NHR¹⁰, —NR¹⁰R¹⁰, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, (C₁-C₈)haloalkyl, —O(C₁-C₈)haloalkyl, —O(C₁-C₈)alkyl,—O(C₁-C₈)alkyleneNHR¹⁰, —O(C₁-C₈)alkyleneNR¹⁰R¹⁰, —(C₁-C₈)alkyleneNHR¹⁰,—(C₁-C₈)alkyleneNR¹⁰R¹⁰, —S(C₁-C₈)alkyl, cycloalkyl, heterocyclyl,heteroaryl, or aryl; R⁷ is —H, —OH, acetyl, —(C₁-C₈)alkyl, —C(O)alkyl,—C(O)cycloalkyl, —C(O)O—(C₁-C₈)alkyl, cycloalkyl, aryl, heteroaryl orheterocyclyl; R^(7a) and R^(7b) independently are —H, —OH, acetyl,—(C₁-C₈)alkyl, —O(C₁-C₈)alkyl, —C(O)alkyl, —C(O)cycloalkyl,—C(O)O—(C₁-C₈)alkyl, cycloalkyl, aryl, heteroaryl or heterocyclyl;R^(8a) and R^(8b) independently are —H, —OH, —CN, acetyl, —SH,—S(O)₂R¹⁰, halogen, —S(C₁-C₈)alkyl, —NHR¹⁰, —NR¹⁰R¹⁰, (C₁-C₈)alkyl,(C₁-C₈)haloalkyl, —O(C₁-C₈)alkyl, —O(C₁-C₈)alkylNHR¹⁰,—O(C₁-C₈)alkylNR¹⁰R¹⁰, —(C₁-C₈)alkylNHR¹⁰, —(C₁-C₈)alkyl NR¹⁰R¹⁰,cycloalkyl, heterocyclyl, heteroaryl or aryl; R⁹, R^(9a) and R^(9b) areindependently —H, (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl,cycloalkyl, heterocyclyl, heteroaryl, aryl, arylalkylene,cycloalkylalkylene, heterocyclylalkylene, or heteroarylalkylene, orR^(9a) and R^(9b) together with the carbon atom to which they areattached form a cycloalkyl or heterocyclyl ring; R¹⁰ is —H, —OH,—C(O)O(C₁-C₈)alkyl, —C(O)(C₁-C₈)alkyl, —C(O)—NH₂, —C(O)—NH(C₁-C₈)alkyl,NH₂—C(O)-alkylene, —S(C₁-C₈)alkyl, acetyl, —(C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, —O(C₁-C₈)alkyl, (C₁-C₈) haloalkyl,alkylcarbonylaminyl, alkylaminyl, —C(O)alkyl, —C(O)cycloalkyl,—C(O)O—(C₁-C₈)alkyl, aryl, heteroaryl, heterocyclyl or cycloalkyl;wherein any alkyl, cycloalkyl, heterocyclyl, heteroaryl, aryl,arylalkylene, cycloalkylalkylene, heterocyclylalkylene,heteroarylalkylene, alkylaminyl, alkylcarbonylaminyl,cycloalkylcarbonylaminyl, cycloalkylaminyl or heterocyclylaminyl isoptionally substituted with 1, 2 or 3 groups selected from —OH, —CN,—SH, —S(O)NH₂, —S(O)NH₂, halogen, —NH₂, —NH(C₁-C₄)alkyl,—N[(C₁-C₄)alkyl]2, —C(O)NH₂, —COOH, —COOMe, acetyl, —(C₁-C₈)alkyl,—O(C₁-C₈)alkyl (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, haloalkyl, thioalkyl,cyanomethylene, alkylaminyl, NH₂—C(O)-alkylene, NH₂—C(O)-alkylene,—NH(Me)-C(O)-alkylene, —CH₂—C(O)-lower alkyl, —C(O)-lower alkyl,alkylcarbonylaminyl, cycloalkyl, cycloalkylalkylene,cycloalkylalkenylene, cycloalkylcarbonylaminyl, cycloalkylaminyl,—CH₂—C(O)-cycloalkyl, —C(O)-cycloalkyl, —CH₂—C(O)-aryl, —CH₂-aryl,—C(O)-aryl, —CH₂—C(O)-heterocycloalkyl, —C(O)-heterocycloalkyl,heterocyclylaminyl or heterocyclyl; and --- represents the option ofhaving a double bond.
 2. The compound according to claim 1 wherein “n”is 1, A¹ is —NR¹⁰, W¹ is O and R¹ is —H.
 3. The compound according toclaim 1 wherein R¹ and R² independently are —H, methyl, ethyl, i-propyl,—NH₂, aminomethylene or —CH₃O—C(O)NH-methylene.
 4. The compoundaccording to claim 1 wherein R¹ and R² together with the carbon atom towhich they are attached form a cycloalkyl ring selected from the groupconsisting of cyclobutyl, cyclopentyl, cyclohexyl,1-(2,2-difluoroethyl)piperidine or 1-methylpiperidine.
 5. The compoundaccording to claim 1 wherein A² is —CR^(5a), A³ is —CR⁶ and A⁴ is—CR^(5b).
 6. The compound according to claim 5 wherein R^(5a) and R⁶independently are —H, chlorine, fluorine, or methyl and R^(5b) is —H. 7.The compound according to claim 1 wherein A² is —CR^(5a), A³ is —CR⁶, A⁴is —N and R^(5a) and R⁶ independently are —H, chlorine, fluorine ormethyl.
 8. The compound according to claim 1 wherein A² is —N, A³ is—CR⁶ and A⁴ is —CR^(5b).
 9. The compound according to claim 8 wherein R⁶is methyl or —H and R^(5b) is —H.
 10. The compound according to claim 1wherein R³ and R⁴ independently are —H.
 11. The compound according toclaim 1 wherein R³ is —H and R⁴ is methyl, ethyl or —NH₂.
 12. Thecompound according to claim 1 wherein --- represents a bond and R^(8a)is —H, heteroaryl or aryl.
 13. The compound according to claim 12wherein A⁶ and A¹ are —CH.
 14. The compound according to claim 12wherein A⁶ is —CH and A⁷ is —C(heteroaryl) or —C(aryl).
 15. The compoundaccording to claim 1 selected from the following table:


16. A pharmaceutical composition comprising (i) a therapeuticallyeffective amount of at least one compound according to claim 1 or astereoisomer, a tautomer or a pharmaceutically acceptable salt thereof;(ii) in combination with a pharmaceutically acceptable carrier, diluentor excipient.
 17. A method for attenuating or inhibiting the activity ofMnK in at least one cell overexpressing Mnk comprising contacting the atleast one cell with a compound according to claim 1 or a stereoisomer,tautomer or pharmaceutically acceptable salt thereof.
 18. The method ofclaim 17 wherein the at least one cell is a colon cancer cell, a gastriccancer cell, a thyroid cancer cell, a lung cancer cell, a leukemia cell,a B-cell lymphoma, a T-cell lymphoma, a hairy cell lymphoma, Hodgkin'slymphoma cell, non-Hodgkin's lymphoma cell, Burkitt's lymphoma cell, apancreatic cancer cell, a melanoma cell, a multiple melanoma cell, abrain cancer cell, a CNS cancer cell, a renal cancer cell, a prostatecancer cell, an ovarian cancer cell, or a breast cancer cell.
 19. Amethod for treating a Mnk dependent condition in a mammal in needthereof comprising administering to the mammal (i) a therapeuticallyeffective amount of at least one compound according to claim 1 or astereoisomer, tautomer or pharmaceutically acceptable salt thereof, or(ii) a pharmaceutical composition of claim
 16. 20. The method of claim19 wherein the Mnk dependent condition is colon cancer, colorectalcancer, gastric cancer, thyroid cancer, lung cancer, leukemia, B-celllymphoma, T-cell lymphoma, hairy cell lymphoma, Hodgkin's lymphoma,non-Hodgkin's lymphoma, Burkitt's lymphoma, pancreatic cancer, melanoma,multiple melanoma, brain cancer, CNS cancer, renal cancer, prostatecancer, ovarian cancer, breast cancer, Alzheimer's, Parkinson's, FragileX Syndrome and autism disorders.